Novel genes encoding proteins having diagnostic, preventive, therapeutic and other uses

ABSTRACT

The invention provides isolated nucleic acids encoding a variety of proteins having diagnostic, preventive, therapeutic, and other uses. These nucleic and proteins are useful for diagnosis, prevention, and therapy of a number of human and other animal disorders. The invention also provides antisense nucleic acid molecules, expression vectors containing the nucleic acid molecules of the invention, host cells into which the expression vectors have been introduced, and non-human transgenic animals in which a nucleic acid molecule of the invention has been introduced or disrupted. The invention still further provides isolated polypeptides, fusion polypeptides, antigenic peptides and antibodies. Diagnostic, screening, and therapeutic methods utilizing compositions of the invention are also provided. The nucleic acids and polypeptides of the present invention are useful as modulating agents in regulating a variety of cellular processes.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation-in-part of co-pending U.S.patent application Ser. No 09/578,063, filed May 24, 2000, which is acontinuation-in-part of co-pending U.S. patent application Ser. No.09/333,159, filed Jun. 14, 1999.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] Not applicable.

REFERENCE TO A MICROFICHE APPENDIX

[0003] Not applicable.

BACKGROUND OF THE INVENTION

[0004] The molecular bases underlying many human and animalphysiological states (e.g., diseased and homeostatic states of varioustissues) remain unknown. Nonetheless, it is well understood that thesestates result from interactions among the proteins and nucleic acidspresent in the cells of the relevant tissues. In the past, thecomplexity of biological systems overwhelmed the ability ofpractitioners to understand the molecular interactions giving rise tonormal and abnormal physiological states. More recently, though, thetechniques of molecular biology, transgenic and null mutant animalproduction, computational biology, pharmacogenomics, and the like haveenabled practitioners to discern the role and importance of individualgenes and proteins in particular physiological states.

[0005] Knowledge of the sequences and other properties of genes(particularly including the portions of genes encoding proteins) and theproteins encoded thereby enables the practitioner to design and screenagents which will affect, prospectively or retrospectively, thephysiological state of an animal tissue in a favorable way. Suchknowledge also enables the practitioner, by detecting the levels of geneexpression and protein production, to diagnose the current physiologicalstate of a tissue or animal and to predict such physiological states inthe future. This knowledge furthermore enables the practitioner toidentify and design molecules which bind with the polynucleotides andproteins, in vitro, in vivo, or both.

[0006] The present invention provides sequence information forpolynucleotides derived from human and murine genes and for proteinsencoded thereby, and thus enables the practitioner to assess, predict,and affect the physiological state of various human and murine tissues.

BRIEF SUMMARY OF THE INVENTION

[0007] The present invention is based, at least in part, on thediscovery of a variety of human and murine cDNA molecules which encodeproteins which are herein designated TANGO 202, TANGO 234, TANGO 265,TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296. These sevenproteins, fragments thereof, derivatives thereof, and variants thereofare collectively referred to herein as the polypeptides of the inventionor the proteins of the invention. Nucleic acid molecules encodingpolypeptides of the invention are collectively referred to as nucleicacids of the invention.

[0008] The nucleic acids and polypeptides of the present invention areuseful as modulating agents in regulating a variety of cellularprocesses. Accordingly, in one aspect, the present invention providesisolated nucleic acid molecules encoding a polypeptide of the inventionor a biologically active portion thereof. The present invention alsoprovides nucleic acid molecules which are suitable as primers orhybridization probes for the detection of nucleic acids encoding apolypeptide of the invention.

[0009] The invention also features nucleic acid molecules which are atleast 40% (or 50%, 60%, 70%, 80%, 90%, 95%, or 98%) identical to thenucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of acDNA clone deposited with ATCC® as one of Accession numbers 207219,207184, 207228, 207185, 207220, and 207221 (“a cDNA of a clone depositedas one of ATCC® 207219, 207184, 207228, 207185, 207220, and 207221”), ora complement thereof.

[0010] The invention features nucleic acid molecules which include afragment of at least 15 (25, 40, 60, 80, 100, 150, 200, 250, 300, 350,400, 450, 550, 650, 700, 800, 900, 1000, 1200, 1400, 1600, 1800, 2000,2200, 2400, 2600, 2800, 3000, 3500, 4000, 4500, or 4928) consecutivenucleotide residues of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of acDNA of a clone deposited as one of ATCC® 207219, 207184, 207228,207185, 207220, and 207221, or a complement thereof.

[0011] The invention also features nucleic acid molecules which includea nucleotide sequence encoding a protein having an amino acid sequencethat is at least 50% (or 60%, 70%, 80%, 90%, 95%, or 98%) identical tothe amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32,35-44, 47-52, 55-66, 69, and 74, or the amino acid sequence encoded by acDNA of a clone deposited as one of ATCC® 207219, 207184, 207228,207185, 207220, and 207221, or a complement thereof.

[0012] In preferred embodiments, the nucleic acid molecules have thenucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or the nucleotide sequenceof a cDNA of a clone deposited as one of ATCC® 207219, 207184, 207228,207185, 207220, and 207221.

[0013] Also within the invention are nucleic acid molecules which encodea fragment of a polypeptide having the amino acid sequence of any of SEQID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, orthe amino acid sequence encoded by a cDNA of a clone deposited as one ofATCC® 207219, 207184, 207228, 207185, 207220, and 207221, the fragmentincluding at least 8 (10, 15, 20, 25, 30, 40, 50, 75, 100, 125, 150, or200) consecutive amino acids of any of SEQ ID NOs: 3-8, 11-16, 19-24,27-32, 35-44, 47-52, 55-66, 69, and 74, or the amino acid sequenceencoded by a cDNA of a clone deposited as one of ATCC® 207219, 207184,207228, 207185, 207220, and 207221.

[0014] The invention includes nucleic acid molecules which encode anaturally occurring allelic variant of a polypeptide comprising theamino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32,35-44, 47-52, 55-66, 69, and 74, or the amino acid sequence encoded by acDNA of a clone deposited as one of ATCC® 207219, 207184, 207228,207185, 207220, and 207221, wherein the nucleic acid molecule hybridizesunder stringent conditions to a nucleic acid molecule having a nucleicacid sequence encoding any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of acDNA of a clone deposited as one of ATCC® 207219, 207184, 207228,207185, 207220, and 207221, or a complement thereof.

[0015] Also within the invention are isolated polypeptides or proteinshaving an amino acid sequence that is at least about 50%, preferably60%, 75%, 90%, 95%, or 98% identical to the amino acid sequence of anyof SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and74.

[0016] Also within the invention are isolated polypeptides or proteinswhich are encoded by a nucleic acid molecule having a nucleotidesequence that is at least about 40%, preferably 50%, 75%, 85%, or 95%identical the nucleic acid sequence encoding any of SEQ ID NOs: 3-8,11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74, and isolatedpolypeptides or proteins which are encoded by a nucleic acid moleculeconsisting of the nucleotide sequence which hybridizes under stringenthybridization conditions to a nucleic acid molecule having thenucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73.

[0017] Also within the invention are polypeptides which are naturallyoccurring allelic variants of a polypeptide that includes the amino acidsequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52,55-66, 69, and 74, or the amino acid sequence encoded by a cDNA of aclone deposited as one of ATCC® 207219, 207184, 207228, 207185, 207220,and 207221, wherein the polypeptide is encoded by a nucleic acidmolecule which hybridizes under stringent conditions to a nucleic acidmolecule having the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9,10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or acomplement thereof.

[0018] The invention also features nucleic acid molecules that hybridizeunder stringent conditions to a nucleic acid molecule having thenucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, the nucleotide sequence of acDNA of a clone deposited as one of ATCC® 207219, 207184, 207228,207185, 207220, and 207221, or a complement thereof. In otherembodiments, the nucleic acid molecules are at least 15 (25, 40, 60, 80,100, 150, 200, 250, 300, 350, 400, 450, 550, 650, 700, 800, 900, 1000,1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3500, 4000,4500, or 4928) nucleotides in length and hybridize under stringentconditions to a nucleic acid molecule having the nucleotide sequence ofany of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54,67, 68, 72, and 73, the nucleotide sequence of a cDNA of a clonedeposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and207221, or a complement thereof. In some embodiments, the isolatednucleic acid molecules encode a cytoplasmic, transmembrane,extracellular, or other domain of a polypeptide of the invention. Inother embodiments, the invention provides an isolated nucleic acidmolecule which is antisense to the coding strand of a nucleic acid ofthe invention.

[0019] Another aspect of the invention provides vectors, e.g.,recombinant expression vectors, comprising a nucleic acid molecule ofthe invention. In another embodiment, the invention provides isolatedhost cells, e.g., mammalian and non-mammalian cells, containing such avector or a nucleic acid of the invention. The invention also providesmethods for producing a polypeptide of the invention by culturing, in asuitable medium, a host cell of the invention containing a recombinantexpression vector encoding a polypeptide of the invention such that thepolypeptide of the invention is produced.

[0020] Another aspect of this invention features isolated or recombinantproteins and polypeptides of the invention. Preferred proteins andpolypeptides possess at least one biological activity possessed by thecorresponding naturally-occurring human polypeptide. An activity, abiological activity, and a functional activity of a polypeptide of theinvention refers to an activity exerted by a protein or polypeptide ofthe invention on a responsive cell as determined in vivo, or in vitro,according to standard techniques.

[0021] Such activities can be a direct activity, such as an associationwith or an enzymatic activity on a second protein, or an indirectactivity, such as a cellular process (e.g., signaling activity) mediatedby interaction of the protein with a second protein. Such activitiesinclude, by way of example, formation of protein-protein interactionswith proteins of one or more signaling pathways (e.g., with a proteinwith which the naturally-occurring polypeptide interacts); binding witha ligand of the naturally-occurring protein; and binding with anintracellular target of the naturally-occurring protein. Otheractivities include modulation of one or more of cellular proliferation,of cellular differentiation, of chemotaxis, of cellular migration, andof cell death (e.g., apoptosis).

[0022] By way of example, TANGO 202 exhibits the ability to affectgrowth, proliferation, survival, differentiation, and activity of humanhematopoietic cells (e.g., bone marrow stromal cells) and fetal cells.TANGO 202 modulates cellular binding to one or more mediators, modulatesproteolytic activity in vivo, modulates developmental processes, andmodulates cell growth, proliferation, survival, differentiation, andactivity. Thus, TANGO 202 can be used to prevent, diagnose, or treatdisorders relating to aberrant cellular protease activity, inappropriateinteraction (or non-interaction) of cells with mediators, inappropriatedevelopment, and blood and hematopoietic cell-related disorders.Exemplary disorders for which TANGO 202 is useful include immunedisorders, infectious diseases, auto-immune disorders, vascular andcardiovascular disorders, disorders related to mal-expression of growthfactors, cancers, hematological disorders, various cancers, birthdefects, developmental defects, and the like.

[0023] Further by way of example, TANGO 234 exhibits the ability toaffect growth, proliferation, survival, differentiation, and activity ofhuman lung, hematopoietic, and fetal cells and of (e.g., bacterial orfungal) cells and viruses which infect humans. TANGO 234 modulatesgrowth, proliferation, survival, differentiation, and activity of gammadelta T cells, for example. Furthermore, TANGO 234 modulates cholesteroldeposition on human arterial walls, and is involved in uptake andmetabolism of low density lipoprotein and regulation of serumcholesterol levels.

[0024] Thus, TANGO 234 can be used to affect development and persistenceof atherogenesis and arteriosclerosis, as well as other vascular andcardiovascular disorders. Other exemplary disorders for which TANGO 234is useful include immune development disorders and disorders involvinggeneration and persistence of an immune response to bacterial, fungal,and viral infections.

[0025] Still further by way of example, TANGO 265 modulates growth andregeneration of neuronal and epithelial tissues, and guides neuronalaxon development. TANGO 265 is a transmembrane protein which mediatescellular interaction with cells, molecules and structures (e.g.,extracellular matrix) in the extracellular environment. TANGO 265 istherefore involved in growth, organization, and adhesion of tissues andthe cells which constitute those tissues. Furthermore, TANGO 265modulates growth, proliferation, survival, differentiation, and activityof neuronal cells and immune system cells. Thus, TANGO 265 can be used,for example, to prevent, diagnose, or treat disorders characterized byaberrant organization or development of a tissue or organ, for guidingneural axon development, for modulating differentiation of cells of theimmune system, for modulating cytokine production by cells of the immunesystem, for modulating reactivity of cells of the immune system towardcytokines, for modulating initiation and persistence of an inflammatoryresponse, and for modulating proliferation of epithelial cells.

[0026] Yet further by way of example, TANGO 273 protein mediates one ormore physiological responses of cells to bacterial infection, e.g., bymediating one or more of detection of bacteria in a tissue in which itis expressed, movement of cells with relation to sites of bacterialinfection, production of biological molecules which inhibit bacterialinfection, and production of biological molecules which alleviatecellular or other physiological damage wrought by bacterial infection.TANGO 273, a transmembrane protein, is also involved in transmembranesignal transduction, and therefore mediates transmission of signalsbetween the extracellular and intracellular environments of cells. TANGO273 mediates regulation of cell growth and proliferation, endocytosis,activation of respiratory burst, and other physiological processestriggered by transmission of a signal via a protein with which TANGO 273interacts. The compositions and methods of the invention can thereforebe used to prevent, diagnose, and treat disorders involving one or morephysiological activities mediated by TANGO 273 protein. Such disordersinclude, for example, various bone-related disorders such as metabolic,homeostatic, and developmental bone disorders (e.g., osteoporosis,various cancers, skeletal development disorders, bone fragility and thelike), disorders caused by or related to bacterial infection, anddisorders characterized by aberrant transmembrane signal transduction byTANGO 273.

[0027] As an additional example, TANGO 286 protein is involved inlipid-binding physiological processes such as lipid transport,metabolism, serum lipid particle regulation, host anti-microbialdefensive mechanisms, and the like. Thus, the compositions and methodsof the invention can therefore be used to prevent, diagnose, and treatdisorders involving one or more physiological activities mediated byTANGO 286 protein. Such disorders include, for example, lipid transportdisorders, lipid metabolism disorders, obesity, disorders of serum lipidparticle regulation, disorders involving insufficient or inappropriatehost anti-microbial defensive mechanisms, vasculitis, bronchiectasis,LPS-related disorders such as shock, disseminated intravascularcoagulation, anemia, thrombocytopenia, adult respiratory distresssyndrome, renal failure, liver disease, and disorders associated withGram negative bacterial infections, such as bacteremia, endotoxemia,sepsis, and the like.

[0028] Further by way of example, TANGO 294 protein is involved infacilitating absorption and metabolism of fat. Thus, the compositionsand methods of the invention can therefore be used to prevent, diagnose,and treat disorders involving one or more physiological activitiesmediated by TANGO 294 protein. Such disorders include, for example,inadequate expression of gastric/pancreatic lipase, cystic fibrosis,exocrine pancreatic insufficiency, medical treatments which alter fatabsorption, obesity, and the like.

[0029] As another example, INTERCEPT 296 protein is involved inphysiological processes related to disorders of the human lung andesophagus. Thus, the compositions and methods of the invention can beused to prevent, diagnose, and treat these disorders. Such disordersinclude, for example, various cancers, bronchitis, cystic fibrosis,respiratory infections (e.g., influenza, bronchiolitis, pneumonia, andtuberculosis), asthma, emphysema, chronic bronchitis, bronchiectasis,pulmonary edema, pleural effusion, pulmonary embolus, adult and infantrespiratory distress syndromes, heartburn, and gastric reflux esophagealdisease.

[0030] In one embodiment, a polypeptide of the invention has an aminoacid sequence sufficiently identical to an identified domain of apolypeptide of the invention. As used herein, the term “sufficientlyidentical” refers to a first amino acid or nucleotide sequence whichcontains a sufficient or minimum number of identical or equivalent(e.g., with a similar side chain) amino acid residues or nucleotides toa second amino acid or nucleotide sequence such that the first andsecond amino acid or nucleotide sequences have a common structuraldomain and/or common functional activity. For example, amino acid ornucleotide sequences which contain a common structural domain havingabout 65% identity, preferably 75% identity, more preferably 85%, 95%,or 98% identity are defined herein as sufficiently identical.

[0031] In one embodiment, the isolated polypeptide of the inventionlacks both a transmembrane and a cytoplasmic domain. In anotherembodiment, the polypeptide lacks both a transmembrane domain and acytoplasmic domain and is soluble under physiological conditions.

[0032] The polypeptides of the present invention, or biologically activeportions thereof, can be operably linked to a heterologous amino acidsequence to form fusion proteins. The invention further featuresantibody substances that specifically bind a polypeptide of theinvention such as monoclonal or polyclonal antibodies, antibodyfragments, single-chain antibodies, and the like. In addition, thepolypeptides of the invention or biologically active portions thereofcan be incorporated into pharmaceutical compositions, which optionallyinclude pharmaceutically acceptable carriers. These antibody substancescan be made, for example, by providing the polypeptide of the inventionto an immunocompetent vertebrate and thereafter harvesting blood orserum from the vertebrate.

[0033] In another aspect, the present invention provides methods fordetecting the presence of the activity or expression of a polypeptide ofthe invention in a biological sample by contacting the biological samplewith an agent capable of detecting an indicator of activity such thatthe presence of activity is detected in the biological sample.

[0034] In another aspect, the invention provides methods for modulatingactivity of a polypeptide of the invention comprising contacting a cellwith an agent that modulates (inhibits or enhances) the activity orexpression of a polypeptide of the invention such that activity orexpression in the cell is modulated. In one embodiment, the agent is anantibody that specifically binds to a polypeptide of the invention.

[0035] In another embodiment, the agent modulates expression of apolypeptide of the invention by modulating transcription, splicing, ortranslation of an mRNA encoding a polypeptide of the invention. In yetanother embodiment, the agent is a nucleic acid molecule having anucleotide sequence that is antisense with respect to the coding strandof an mRNA encoding a polypeptide of the invention.

[0036] The present invention also provides methods to treat a subjecthaving a disorder characterized by aberrant activity of a polypeptide ofthe invention or aberrant expression of a nucleic acid of the inventionby administering an agent which is a modulator of the activity of apolypeptide of the invention or a modulator of the expression of anucleic acid of the invention to the subject. In one embodiment, themodulator is a protein of the invention. In another embodiment, themodulator is a nucleic acid of the invention. In other embodiments, themodulator is a peptide, peptidominetic, or other small molecule (e.g., asmall organic molecule).

[0037] The present invention also provides diagnostic assays foridentifying the presence or absence of a genetic lesion or mutationcharacterized by at least one of: (i) aberrant modification or mutationof a gene encoding a polypeptide of the invention, (ii) mis-regulationof a gene encoding a polypeptide of the invention, and (iii) aberrantpost-translational modification of a polypeptide of the inventionwherein a wild-type form of the gene encodes a polypeptide having theactivity of the polypeptide of the invention.

[0038] In another aspect, the invention provides a method foridentifying a compound that binds to or modulates the activity of apolypeptide of the invention. In general, such methods entail measuringa biological activity of the polypeptide in the presence and absence ofa test compound and identifying those compounds which alter the activityof the polypeptide.

[0039] The invention also features methods for identifying a compoundwhich modulates the expression of a polypeptide or nucleic acid of theinvention by measuring the expression of the polypeptide or nucleic acidin the presence and absence of the compound.

[0040] In yet a further aspect, the invention provides substantiallypurified antibodies or fragments thereof (i.e., antibody substances),including non-human antibodies or fragments thereof, which specificallybind with a polypeptide of the invention or with a portion thereof. Invarious embodiments, these substantially purified antibodies/fragmentscan be human, non-human, chimeric, and/or humanized antibodies.Non-human antibodies included in the invention include, by way ofexample, goat, mouse, sheep, horse, chicken, rabbit, and rat antibodies.In addition, the antibodies of the invention can be polyclonalantibodies or monoclonal antibodies.

[0041] In a particularly preferred embodiment, the antibody substance ofthe invention specifically binds with an extracellular domain of one ofTANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, andINTERCEPT 296. Preferably, the extracellular domain with which theantibody substance binds has an amino acid sequence selected from thegroup consisting of SEQ ID NOs: 5, 6, 14, 22, 30, 37, 49, 50, and 56-58.

[0042] Any of the antibody substances of the invention can be conjugatedwith a therapeutic moiety or with a detectable substance. Non-limitingexamples of detectable substances that can be conjugated with theantibody substances of the invention include an enzyme, a prostheticgroup, a fluorescent material (i.e., a fluorophore), a luminescentmaterial, a bioluminescent material, and a radioactive material (e.g., aradionuclide or a substituent comprising a radionuclide).

[0043] The invention also provides a kit containing an antibodysubstance of the invention conjugated with a detectable substance, andinstructions for use. Still another aspect of the invention is apharmaceutical composition comprising an antibody substance of theinvention and a pharmaceutically acceptable carrier. In preferredembodiments, the pharmaceutical composition contains an antibodysubstance of the invention, a therapeutic moiety (preferably conjugatedwith the antibody substance), and a pharmaceutically acceptable carrier.

[0044] The invention includes a method of assessing whether a firsthuman patient is afflicted with an epithelial or endothelial tumor(e.g., a colon, prostate, pancreatic, lung, or breast tumor). The methodcomprises comparing:

[0045] a) occurrence of a nucleic acid molecule of claim 1 in a sampleobtained from the first patient and

[0046] b) occurrence of the nucleic acid molecule in a control sampleselected from the group consisting of

[0047] i) a control sample obtained from a tissue that is obtained fromthe first patient and that is known not to comprise the tumor; and

[0048] ii) a control sample obtained from a second patient who is knownnot to be afflicted with the tumor.

[0049] A difference between the first sample and the control sample isan indication that the patient is afflicted with the tumor.

[0050] The invention also includes a method of assessing whether a firsthuman patient is afflicted with an epithelial or endothelial tumor. Thismethod comprises comparing

[0051] a) occurrence of a nucleic acid molecule of claim 1 in a sampleobtained from the first patient and

[0052] b) occurrence of the nucleic acid molecule in a control sampleselected from the group consisting of

[0053] i) a control sample obtained from a tissue that is obtained fromthe first patient and that is known to comprise the tumor; and

[0054] ii) a control sample obtained from a second patient who is knownto be afflicted with the tumor.

[0055] A difference between the first sample and the control sample isan indication that the patient is not afflicted with the tumor.

[0056] In another embodiment, the invention includes a method ofscreening for agents which decrease the activity of a TANGO-294-likelipase protein. The method comprises:

[0057] contacting a test compound with a TANGO 294-like lipasepolypeptide encoded by an isolated nucleic acid molecule of claim 1 and

[0058] detecting binding between the test compound and the TANGO294-like lipase polypeptide,

[0059] Binding between the test compound and the TANGO 294-like lipasepolypeptide is an indication that the test compound is an agent whichdecreases the activity of the TANGO 294-like lipase protein.

[0060] A pharmaceutical composition can be made by identifying an agentaccording to this method and combining the agent and a pharmaceuticallyacceptable carrier to form the pharmaceutical composition. Thispharmaceutical composition can be administered to a human afflicted witha disorder in order to modulate the activity of a TANGO 294-like lipaseprotein in the disorder. The disorder can, for example be a tumor, adisorder of fat absorption, a disorder of fat metabolism, a blood flowdisorder, a blood pressure disorder, an inflammatory disorder, an immunedisorder, a thrombotic disorder, or a disorder involving inappropriateplatelet adherence. Specific examples of these disorders include tumorsof endothelial or epithelial origin, colon tumors, pancreatic tumors,inadequate expression of gastric lipase, inadequate expression ofpancreatic lipase, cystic fibrosis, exocrine pancreatic insufficiency,obesity, arterial hypertension, renovascular hypertension, syncope,orthostatic hypotension, shock, gastritis, gastric ulcer, colitis,irritable bowel syndrome, inflammatory bowel syndrome, dermatitis,pancreatitis, rheumatoid arthritis, psoriasis, myasthenia gravis, anallergy, insulin resistance, systemic lupus erythematosus, scleroderma,and autoimmune diabetes mellitus, an infection of a human by aninfectious agent (e.g., human immunodeficiency virus), hemophilia,stroke, myocardial infarction, coronary artery disease, andatherosclerosis.

[0061] In still another embodiment, the invention includes a method ofscreening for agents which modulate the activity of a TANGO 294-likelipase protein. This method comprises:

[0062] contacting a test compound with a TANGO 294-like lipasepolypeptide encoded by an isolated nucleic acid molecule of claim 1 and

[0063] detecting a TANGO 294-like lipase activity of the polypeptide,

[0064] Increased TANGO 294-like lipase activity in the presence of thetest compound is an indication that the test compound is an agent usefulfor increasing the activity of the TANGO 294-like lipase protein.Decreased TANGO 294-like lipase activity in the presence of the testcompound is an indication that the test compound is an agent useful fordecreasing the activity of the TANGO 294-like lipase protein.

[0065] A pharmaceutical composition can be made by identifying an agentaccording to this method and combining the agent and a pharmaceuticallyacceptable carrier to form the pharmaceutical composition. Thispharmaceutical composition can be administered to a human afflicted witha disorder in order to modulate the activity of a TANGO 294-like lipaseprotein in the disorder. The disorder can, for example be a tumor, adisorder of fat absorption, a disorder of fat metabolism, a blood flowdisorder, a blood pressure disorder, an inflammatory disorder, an immunedisorder, a thrombotic disorder, or a disorder involving inappropriateplatelet adherence. Specific examples of these disorders include tumorsof endothelial or epithelial origin, colon tumors, pancreatic tumors,inadequate expression of gastric lipase, inadequate expression ofpancreatic lipase, cystic fibrosis, exocrine pancreatic insufficiency,obesity, arterial hypertension, renovascular hypertension, syncope,orthostatic hypotension, shock, gastritis, gastric ulcer, colitis,irritable bowel syndrome, inflammatory bowel syndrome, dermatitis,pancreatitis, rheumatoid arthritis, psoriasis, myasthenia gravis, anallergy, insulin resistance, systemic lupus erythematosus, scleroderna,and autoimmune diabetes mellitus, an infection of a human by aninfectious agent (e.g., human immunodeficiency virus), hemophilia,stroke, myocardial infarction, coronary artery disease, andatherosclerosis.

[0066] In yet another embodiment, the invention includes a method ofscreening for agents which decrease the activity of a TANGO 294-likelipase protein. This method comprises:

[0067] contacting a test compound with an isolated nucleic acid moleculeof claim 1 and

[0068] detecting binding of the test compound with the isolated nucleicacid molecule,

[0069] Binding between the test compound and the isolated nucleic acidmolecule is an indication that the test compound is a agent useful fordecreasing the activity of the TANGO 294-like lipase protein.

[0070] A pharmaceutical composition can be made by identifying an agentaccording to this method and combining the agent and a pharmaceuticallyacceptable carrier to form the pharmaceutical composition. Thispharmaceutical composition can be administered to a human afflicted witha disorder in order to modulate the activity of a TANGO 294-like lipaseprotein in the disorder. The disorder can, for example be a tumor, adisorder of fat absorption, a disorder of fat metabolism, a blood flowdisorder, a blood pressure disorder, an inflammatory disorder, an immunedisorder, a thrombotic disorder, or a disorder involving inappropriateplatelet adherence. Specific examples of these disorders include tumorsof endothelial or epithelial origin, colon tumors, pancreatic tumors,inadequate expression of gastric lipase, inadequate expression ofpancreatic lipase, cystic fibrosis, exocrine pancreatic insufficiency,obesity, arterial hypertension, renovascular hypertension, syncope,orthostatic hypotension, shock, gastritis, gastric ulcer, colitis,irritable bowel syndrome, inflammatory bowel syndrome, dermatitis,pancreatitis, rheumatoid arthritis, psoriasis, myasthenia gravis, anallergy, insulin resistance, systemic lupus erythematosus, scleroderma,and autoimmune diabetes mellitus, an infection of a human by aninfectious agent (e.g., human immunodeficiency virus), hemophilia,stroke, myocardial infarction, coronary artery disease, andatherosclerosis.

[0071] The invention also includes a method of reducing the activity ofa TANGO 294-like lipase protein (i.e., TANGO 294, or a variant oralternative form thereof as disclosed herein) of a cell. The methodcomprises contacting the cell with a reagent which specifically bindswith an isolated TANGO 294 nucleic acid molecule or with an isolatedTANGO 294 polypeptide.

[0072] Other features and advantages of the invention will be apparentfrom the following detailed description and claims.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0073]FIG. 1 comprises FIGS. 1A-1M. The nucleotide sequence (SEQ IDNO: 1) of a cDNA encoding the human TANGO 202 protein described hereinis listed in FIGS 1A-1D. The open reading frame (ORF; residues 34 to1458; SEQ ID NO: 2) of the cDNA is indicated by nucleotide triplets,above which the amino acid sequence (SEQ ID NO: 3) of human TANGO 202 islisted. The nucleotide sequence (SEQ ID NO: 67) of a cDNA encoding themurine TANGO 202 protein described herein is listed in FIGS. 1E-1I. TheORF (residues 81 to 1490; SEQ ID NO: 68) of the cDNA is indicated bynucleotide triplets, above which the amino acid sequence (SEQ ID NO: 69)of murine TANGO 202 is listed. An alignment of the amino acid sequencesof human (“Hum.”; SEQ ID NO: 3) and murine (“Mur.”; SEQ ID NO: 69) TANGO202 protein is shown in FIGS. 1J and 1K, wherein identical amino acidresidues are indicated by “:” and similar amino acid residues areindicated by “.”.

[0074]FIG. 1L is a hydrophilicity plot of human TANGO 202 protein, inwhich the locations of cysteine residues (“Cys”) and potentialN-glycosylation sites (“Ngly”) are indicated by vertical bars and thepredicted extracellular (“out”), intracellular (“ins”), or transmembrane(“TM”) locations of the protein backbone is indicated by a horizontalbar.

[0075]FIG. 1M is a hydrophilicity plot of murine TANGO 202 protein.

[0076]FIG. 2 comprises FIGS. 2A to 2Q-17. The nucleotide sequence (SEQID NO: 9) of a cDNA encoding the human TANGO 234 protein describedherein is listed in FIGS. 2A-2I. The ORF (residues 28 to 4386; SEQ IDNO: 10) of the cDNA is indicated by nucleotide triplets, above which theamino acid sequence (SEQ ID NO: 11) of human TANGO 234 is listed.

[0077]FIG. 2J is a hydrophilicity plot of human TANGO 234 protein. Analignment of the amino acid sequences of human TANGO 234 (“Hum”; SEQ IDNO: 11) and bovine WC1 (“WC1”; SEQ ID NO: 78) proteins is shown in FIGS.2K-2P, wherein identical amino acid residues are indicated by “:” andsimilar amino acid residues are indicated by “.”. An alignment of thenucleotide sequences of an ORF encoding human TANGO 234 (“Hum”; SEQ IDNO: 10) and an ORF encoding bovine WC1 (“WC1”; SEQ ID NO: 79) proteinsis shown in FIGS. 2Q-1 to 2Q-17, wherein identical nucleotide residuesare indicated by “:”.

[0078]FIG. 3 comprises FIGS. 3A-3U. The nucleotide sequence (SEQ ID NO:17) of a cDNA encoding the human TANGO 265 protein described herein islisted in FIGS. 3A-3E. The ORF (residues 32 to 2314; SEQ ID NO: 18) ofthe cDNA is indicated by nucleotide triplets, above which the amino acidsequence (SEQ ID NO: 19) of human TANGO 265 is listed. An alignment ofthe amino acid sequences of human TANGO 265 protein (“Hum.”; SEQ ID NO:19) and murine semaphorin B protein (“Mur.”; SEQ ID NO: 70; GenBankAccession No. X85991) is shown in FIGS. 3F-3H, wherein identical aminoacid residues are indicated by “:” and similar amino acid residues areindicated by “.”.

[0079] In FIGS. 31-3T, an alignment of the nucleotide sequences of thecDNA encoding human TANGO 265 protein (“Hum.”; SEQ ID NO: 17) and thenucleotide sequences of the cDNA encoding murine semaphorin B protein(“Mur.”; SEQ ID NO: 71; GenBank Accession No. X85991) is shown.

[0080]FIG. 3U is a hydrophilicity plot of TANGO 265 protein.

[0081]FIG. 4 comprises FIGS. 4A-4J. The nucleotide sequence (SEQ ID NO:25) of a cDNA encoding the human TANGO 273 protein described herein islisted in FIGS. 4A-4C. The ORF (residues 135 to 650; SEQ ID NO: 26) ofthe cDNA is indicated by nucleotide triplets, above which the amino acidsequence (SEQ ID NO: 27) of human TANGO 273 is listed. The nucleotidesequence (SEQ ID NO: 72) of a cDNA encoding the murine TANGO 273 proteindescribed herein is listed in FIGS. 4D-4G. The ORF (residues 137 to 652;SEQ ID NO: 73) of the cDNA is indicated by nucleotide triplets, abovewhich the amino acid sequence (SEQ ID NO: 74) of murine TANGO 273 islisted. An alignment of the amino acid sequences of human (“Hum.”; SEQID NO: 27) and murine (“Mur.”; SEQ ID NO: 74) TANGO 273 protein is shownin FIG. 4H, wherein identical amino acid residues are indicated by “:”and similar amino acid residues are indicated by “.”.

[0082]FIG. 4J is a hydrophilicity plot of human TANGO 273 protein, and

[0083]FIG. 4J is a hydrophilicity plot of murine TANGO 273 protein.

[0084]FIG. 5 comprises FIGS. 5A-5I. The nucleotide sequence (SEQ ID NO:33) of a cDNA encoding the human TANGO 286 protein described herein islisted in FIGS. 5A-5D. The ORF (residues 133 to 1497; SEQ ID NO: 34) ofthe cDNA is indicated by nucleotide triplets, above which the amino acidsequence (SEQ ID NO: 35) of human TANGO 286 is listed.

[0085]FIG. 5E is a hydrophilicity plot of TANGO 286 protein. Analignment of the amino acid sequences of human TANGO 286 (“286”; SEQ IDNO: 35) and BPI protein (“BPI”; SEQ ID NO: 38) protein is shown in FIGS.5F and 5G, wherein identical amino acid residues are indicated by “:”and similar amino acid residues are indicated by “.”. An alignment ofthe amino acid sequences of human TANGO 286 (“286”; SEQ ID NO: 35) andRENP protein (“RENP”; SEQ ID NO: 39) is shown in FIGS. 5H and 5I,wherein identical amino acid residues are indicated by “:” and similaramino acid residues are indicated by “.”.

[0086]FIG. 6 comprises FIGS. 6A-6H. The nucleotide sequence (SEQ ID NO:45) of a cDNA encoding the human TANGO 294 protein described herein islisted in FIGS. 6A-6C. The ORF (residues 126 to 1394; SEQ ID NO: 46) ofthe cDNA is indicated by nucleotide triplets, above which the amino acidsequence (SEQ ID NO: 47) of human TANGO 294 is listed. An alignment ofthe amino acid sequences of human TANGO 294 protein (“294”; SEQ ID NO:47) and a known human lipase protein (“HLP”; SEQ ID NO: 75; GenBankAccession No. NP_(—)004181) is shown in FIGS. 6D and 6E, whereinidentical amino acid residues are indicated by “:” and similar aminoacid residues are indicated by “.”.

[0087]FIG. 6F is a hydrophilicity plot of TANGO 294 protein. Analignment of the amino acid sequences of human TANGO 294 protein (“294”;SEQ ID NO: 47) and a known human lysosomal acid lipase protein (“LAL”;SEQ ID NO: 41) is shown in FIGS. 6G and 6H, wherein identical amino acidresidues are indicated by and similar amino acid residues are indicatedby “.”

[0088]FIG. 7 comprises FIGS. 7A-7F. The nucleotide sequence (SEQ ID NO:53) of a cDNA encoding the human INTERCEPT 296 protein described hereinis listed in FIGS. 7A-7C. The ORF (residues 70 to 1098; SEQ ID NO: 54)of the cDNA is indicated by nucleotide triplets, above which the aminoacid sequence (SEQ ID NO: 55) of human INTERCEPT 296 protein is listed.

[0089]FIG. 7D is a hydrophilicity plot of INTERCEPT 296 protein. Analignment of the amino acid sequences of human INTERCEPT 296 protein(“296”; SEQ ID NO: 55) and C. elegans C06E1.3 related protein (“CRP”;SEQ ID NO: 40) is shown in FIGS. 7E and 7F, wherein identical amino acidresidues are indicated by “:” and similar amino acid residues areindicated by “.”.

DETAILED DESCRIPTION OF THE INVENTION

[0090] The present invention is based, at least in part, on thediscovery of a variety of human and murine cDNA molecules which encodeproteins which are herein designated TANGO 202, TANGO 234, TANGO 265,TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296. These proteinsexhibit a variety of physiological activities, and are included in asingle application for the sake of convenience. It is understood thatthe allowability or non-allowability of claims directed to one of theseproteins has no bearing on the allowability of claims directed to theothers. The characteristics of each of these proteins and the cDNAsencoding them are now described separately.

[0091] TANGO 202

[0092] A cDNA clone (designated jthke096b05) encoding at least a portionof human TANGO 202 protein was isolated from a human fetal skin cDNAlibrary. The corresponding murine cDNA was isolated as a clone(designated jtmMa044f07) from a bone marrow stromal cell cDNA library.The human TANGO 202 protein is predicted by structural analysis to be atype I membrane protein, although it can exist in a secreted form aswell. The murine TANGO 202 protein is predicted by structural analysisto be a secreted protein.

[0093] The full length of the cDNA encoding human TANGO 202 protein(FIG. 1; SEQ ID NO: 1) is 1656 nucleotide residues. The open readingframe (ORF) of this cDNA, nucleotide residues 34 to 1458 of SEQ ID NO: 1(i.e., SEQ ID NO: 2), encodes a 475-amino acid transmembrane protein(FIG. 1; SEQ ID NO: 3).

[0094] The invention thus includes purified human TANGO 202 protein,both in the form of the immature 475 amino acid residue protein (SEQ IDNO: 3) and in the form of the mature 456 amino acid residue protein (SEQID NO: 5). The invention also includes purified murine TANGO 202protein, both in the form of the immature 470 amino acid residue protein(SEQ ID NO: 67) and in the form of the mature 451 amino acid residueprotein (SEQ ID NO: 43). Mature human or murine TANGO 202 proteins canbe synthesized without the signal sequence polypeptide at the aminoterminus thereof, or they can be synthesized by generating immatureTANGO 202 protein and cleaving the signal sequence therefrom.

[0095] In addition to full length mature and immature human and murineTANGO 202 proteins, the invention includes fragments, derivatives, andvariants of these TANGO 202 proteins, as described herein. Theseproteins, fragments, derivatives, and variants are collectively referredto herein as polypeptides of the invention or proteins of the invention.

[0096] The invention also includes nucleic acid molecules which encode apolypeptide of the invention. Such nucleic acids include, for example, aDNA molecule having the nucleotide sequence listed in SEQ ID NO: 1 orsome portion thereof or SEQ ID NO: 67 or some portion thereof, such asthe portion which encodes mature human or murine TANGO 202 protein,immature human or murine TANGO 202 protein, or a domain of human ormurine TANGO 202 protein. These nucleic acids are collectively referredto as nucleic acids of the invention.

[0097] TANGO 202 proteins and nucleic acid molecules encoding themcomprise a family of molecules having certain conserved structural andfunctional features. As used herein, the term “family” is intended tomean two or more proteins or nucleic acid molecules having a common orsimilar domain structure and having sufficient amino acid or nucleotidesequence identity as defined herein. Family members can be from eitherthe same or different species (e.g., human and mouse, as describedherein). For example, a family can comprise two or more proteins ofhuman origin, or can comprise one or more proteins of human origin andone or more of non-human origin.

[0098] A common domain present in TANGO 202 proteins is a signalsequence. As used herein, a signal sequence includes a peptide of atleast about 10 amino acid residues in length which occurs at the aminoterminus of membrane-bound and secreted proteins and which contains atleast about 45% hydrophobic amino acid residues such as alanine,leucine, isoleucine, phenylalanine, proline, tyrosine, tryptophan, orvaline. In a preferred embodiment, a signal sequence contains at leastabout 10 to 35 amino acid residues, preferably about 10 to 20 amino acidresidues, and has at least about 35-60%, more preferably 40-50%, andmore preferably at least about 45% hydrophobic residues. A signalsequence serves to direct a protein containing such a sequence to alipid bilayer. Thus, in one embodiment, a TANGO 202 protein contains asignal sequence corresponding to amino acid residues 1 to 19 of SEQ IDNO: 3 (SEQ ID NO: 4) or to amino acid residues 1 to 19 of SEQ ID NO: 69(SEQ ID NO: 42). The signal sequence is cleaved during processing of themature protein.

[0099] TANGO 202 proteins can also include an extracellular domain. Asused herein, an “extracellular domain” refers to a portion of a proteinwhich is localized to the non-cytoplasmic side of a lipid bilayer of acell when a nucleic acid encoding the protein is expressed in the cell.The human TANGO 202 protein extracellular domain is located from aboutamino acid residue 20 to about amino acid residue 392 of SEQ ID NO: 3 inthe non-secreted form, and from about amino acid residue 20 to aminoacid residue 475 of SEQ ID NO: 3 (i.e., the entire mature humanprotein). The murine TANGO 202 protein extracellular domain is locatedfrom about amino acid residue 20 to amino acid residue 470 of SEQ ID NO:69 (i.e., the entire mature murine protein).

[0100] TANGO 202 proteins of the invention can also include atransmembrane domain. As used herein, a “transmembrane domain” refers toan amino acid sequence having at least about 20 to 25 amino acidresidues in length and which contains at least about 65-70% hydrophobicamino acid residues such as alanine, leucine, phenylalanine, protein,tyrosine, tryptophan, or valine. In a preferred embodiment, atransmembrane domain contains at least about 15 to 30 amino acidresidues, preferably about 20-25 amino acid residues, and has at leastabout 60-80%, more preferably 65-75%, and more preferably at least about70% hydrophobic residues. Thus, in one embodiment, a TANGO 202 proteinof the invention contains a transmembrane domain corresponding to aboutamino acid residues 393 to 415 of SEQ ID NO: 3 (SEQ ID NO: 7).

[0101] In addition, TANGO 202 proteins of the invention can include acytoplasmic domain, particularly including a carboxyl-terminalcytoplasmic domain. As used herein, a “cytoplasmic domain” refers to aportion of a protein which is localized to the cytoplasmic side of alipid bilayer of a cell when a nucleic acid encoding the protein isexpressed in the cell. The cytoplasmic domain is located from aboutamino acid residue 416 to amino acid residue 475 of SEQ ID NO: 3 (SEQ IDNO: 8) in the non-secreted form of human TANGO 202 protein.

[0102] TANGO 202 proteins typically comprise a variety of potentialpost-translational modification sites (often within an extracellulardomain), such as those described herein in Tables I (for human TANGO202) and II (for murine TANGO 202), as predicted by computerizedsequence analysis of TANGO 202 proteins using amino acid sequencecomparison software (comparing the amino acid sequence of TANGO 202 withthe information in the PROSITE database {rel. 12.2; February, 1995} andthe Hidden Markov Models database {Rel. PFAM 3.3}). TABLE I Amino AcidType of Potential Modification Site or Residues of Amino Acid Domain SEQID NO: 3 Sequence N-glycosylation site 47 to 50 NWTA 61 to 64 NETF 219to 222 NYSA 295 to 298 NVSL 335 to 338 NQTV 347 to 350 NLSV Proteinkinase C phosphorylation site 70 to 72 TLK 137 to 139 TSK 141 to 143 SNK155 to 157 SQR 238 to 240 TGR 245 to 247 TIR 277 to 279 THR 307 to 309SDR 355 to 357 SSK 387 to 389 SHR 418 to 420 TFK 421 to 423 SHR Caseinkinase II phosphorylation site 337 to 340 TVAE 438 to 441 TSGE 464 to467 SQQD N-myristoylation site 53 to 58 GGKPCL 120 to 125 GNLGCY 136 to141 GTSKTS 162 to 167 GMESGY 214 to 219 GACGGN Kringle domain signature85 to 90 YCRNPD Kringle Domain  34 to 116 See FIG. 1 CUB domain 216 to320 See FIG. 1

[0103] TABLE II Amino Acid Type of Potential Modification Residues ofAmino Acid Site or Domain SEQ ID NO: 69 Sequence N-glycosylation site 59to 62 NETF 217 to 220 NYSA 255 to 258 NFTL 293 to 296 NVSL 333 to 336NQTL 345 to 348 NLSV cAMP- or cGMP-dependent protein 455 to 458 RRSSkinase phosphorylation site Protein kinase C phosphorylation site 68 to70 TLK 135 to 137 TSK 139 to 141 SNK 153 to 155 SQR 236 to 238 TGR 243to 245 TIR 275 to 277 THR 283 to 285 SGR 305 to 307 SDR 353 to 355 SSK408 to 410 SQR 453 to 455 SLR 457 to 459 SSR Casein kinase IIphosphorylation site 28 to 31 SGPE 257 to 260 TLFD 321 to 324 TKEE 335to 338 TLAE 384 to 387 TATE N-myristoylation site 51 TO 56 GGKPCL 118 TO123 GNLGCY 134 TO 139 GTSKTS 160 TO 165 GMESGY 212 TO 217 GACGGN 391 TO396 GLCTAW 429 TO 434 GTVVSL Kringle domain signature 83 to 88 YCRNPDKringle Domain  32 to 114 See FIG. 1 CUB domain 214 to 318 See FIG. 1

[0104] As used herein, the term “post-translational modification site”refers to a protein domain that includes about 3 to 10 amino acidresidues, more preferably about 3 to 6 amino acid residues wherein thedomain has an amino acid sequence which comprises a consensus sequencewhich is recognized and modified by a protein-modifying enzyme.Exemplary protein-modifying enzymes include amino acid glycosylases,cAMP- and cGMP-dependent protein kinases, protein kinase C, caseinkinase II, myristoylases, and prenyl transferases. In variousembodiments, the protein of the invention has at least 1, 2, 4, 6, 10,15, or 20 or more of the post-translational modification sites describedherein in Tables I and II.

[0105] Exemplary additional domains present in human and murine TANGO202 protein include Kringle domains and CUB domains. In one embodiment,the protein of the invention has at least one domain that is at least55%, preferably at least about 65%, more preferably at least about 75%,yet more preferably at least about 85%, and most preferably at leastabout 95% identical to one of the domains described herein in Tables Iand II. Preferably, the protein of the invention has at least oneKringle domain and one CUB domain.

[0106] A Kringle domain has a characteristic profile that has beendescribed in the art (Castellino and Beals (1987) J. Mol. Evol.26:358-369; Patthy (1985) Cell 41:657-663; Ikeo et al. (1991) FEBS Lett.287:146-148). Many, but not all, Kringle domains comprise a conservedhexapeptide signature sequence, namely

(F or Y)-C-R-N-P-(D or N or R).

[0107] The cysteine residue is involved in a disulfide bond.

[0108] Kringle domains are triple-looped, disulfide cross-linked domainsfound in a varying number of copies in, for example, some serineproteases and plasma proteins. Kringle domains have a role in bindingmediators (e.g., membranes, other proteins, or phospholipids) and inregulation of proteolytic activity. Kringle domains have been identifiedin the following proteins, for example: apolipoprotein A, bloodcoagulation factor XII (Hageman factor), hepatocyte growth factor (HGF),HGF-like protein (Friezner Degen et al., (1991) Biochemistry30:9781-9791), HGF activator (Miyazawa et al., (1993) J. Biol. Chem.268:10024-10028), plasminogen, thrombin, tissue plasminogen activator,urokinase-type plasminogen activator, and four influenza neuraminidases.The presence of a Kringle domain in each of human and murine TANGO 202protein indicates that TANGO 202 is involved in one or morephysiological processes in which these other Kringle domain-containingproteins are involved, has biological activity in common with one ormore of these other Kringle domain-containing proteins, or both.

[0109] CUB domains are extracellular domains of about 110 amino acidresidues which occur in functionally diverse, mostly developmentallyregulated proteins (Bork and Beckmann (1993) J. Mol. Biol. 231:539-545;Bork (1991) FEBS Lett. 282:9-12). Many CUB domains contain fourconserved cysteine residues, although some, like that of TANGO 202,contain only two of the conserved cysteine residues. The structure ofthe CUB domain has been predicted to assume a beta-barrel configuration,similar to that of immunoglobulins. Other proteins which have been foundto comprise one or more CUB domains include, for example, mammaliancomplement sub-components Cls and Clr, hamster serine protease Casp,mammalian complement activating component of Ra-reactive factor,vertebrate enteropeptidase, vertebrate bone morphogenic protein 1, seaurchin blastula proteins BP10 and SpAN, Caenorhabditis eleganshypothetical proteins F42A10.8 and R151.5, neuropilin (A5 antigen), seaurchin fibropellins I and III, mammalian hyaluronate-binding proteinTSG-6 (PS4), mammalian spermadhesins, and Xenopus embryonic proteinUVS.2. The presence of a CUB domain in each of human and murine TANGO202 protein indicates that TANGO 202 is involved in one or morephysiological processes in which these other CUB domain-containingproteins are involved, has biological activity in common with one ormore of these other CUB domain-containing proteins, or both.

[0110] The signal peptide prediction program SIGNALP (Nielsen et al.(1997) Protein Engineering 10:1-6) predicted that human TANGO 202protein includes a 19 amino acid signal peptide (amino acid residues 1to 19 of SEQ ID NO: 3; SEQ ID NO: 4) preceding the mature TANGO 202protein (amino acid residues 20 to 475 of SEQ ID NO: 3; SEQ ID NO: 5).Human TANGO 202 protein includes an extracellular domain (amino acidresidues 20 to 392 of SEQ ID NO: 3; SEQ ID NO: 6); a transmembranedomain (amino acid residues 393 to 415 of SEQ ID NO: 3; SEQ ID NO: 7);and a cytoplasmic domain (amino acid residues 416 to 475 of SEQ ID NO:3; SEQ ID NO: 8). The murine homolog of TANGO 202 protein is predictedto be a secreted protein. Thus, it is recognized that human TANGO 202can also exist in the form of a secreted protein, likely beingtranslated from an alternatively spliced TANGO 202 mRNA. In a variantform of the protein, an extracellular portion of TANGO 202 protein(e.g., amino acid residues 20 to 392 of SEQ ID NO: 3) can be cleavedfrom the mature protein to generate a soluble fragment of TANGO 202.

[0111]FIG. 1L depicts a hydrophilicity plot of human TANGO 202 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Thehydrophobic region which corresponds to amino acid residues 1 to 19 ofSEQ ID NO: 3 is the signal sequence of human TANGO 202 (SEQ ID NO: 4).The hydrophobic region which corresponds to amino acid residues 393 to415 of SEQ ID NO: 3 is the transmembrane domain of human TANGO 202 (SEQID NO: 7). As described elsewhere herein, relatively hydrophilic regionsare generally located at or near the surface of a protein, and are morefrequently effective immunogenic epitopes than are relativelyhydrophobic regions. For example, the region of human TANGO 202 proteinfrom about amino acid residue 61 to about amino acid residue 95 appearsto be located at or near the surface of the protein, while the regionfrom about amino acid residue 395 to about amino acid residue 420appears not to be located at or near the surface.

[0112] The predicted molecular weight of human TANGO 202 protein withoutmodification and prior to cleavage of the signal sequence is about 51.9kilodaltons. The predicted molecular weight of the mature human TANGO202 protein without modification and after cleavage of the signalsequence is about 50.1 kilodaltons.

[0113] The full length of the cDNA encoding murine TANGO 202 protein(FIG. 1; SEQ ID NO: 67) is 4928 nucleotide residues. The ORF of thiscDNA, nucleotide residues 81 to 1490 of SEQ ID NO: 67 (i.e., SEQ ID NO:68), encodes a 470-amino acid secreted protein (FIG. 1; SEQ ID NO: 69).

[0114] The signal peptide prediction program SIGNALP (Nielsen et al.(1997) Protein Engineering 10:1-6) predicted that murine TANGO 202protein includes a 19 amino acid signal peptide (amino acid residues 1to 19 of SEQ ID NO: 69; SEQ ID NO: 42) preceding the mature TANGO 202protein (amino acid residues 20 to 470 of SEQ ID NO: 69; SEQ ID NO: 43).Murine TANGO 202 protein is a secreted protein.

[0115]FIG. 1M depicts a hydrophilicity plot of murine TANGO 202 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Thehydrophobic region which corresponds to amino acid residues 1 to 19 ofSEQ ID NO: 69 is the signal sequence of murine TANGO 202 (SEQ ID NO:42). As described elsewhere herein, relatively hydrophilic regions aregenerally located at or near the surface of a protein, and are morefrequently effective immunogenic epitopes than are relativelyhydrophobic regions. For example, the region of murine TANGO 202 proteinfrom about amino acid residue 61 to about amino acid residue 95 appearsto be located at or near the surface of the protein, while the regionfrom about amino acid residue 295 to about amino acid residue 305appears not to be located at or near the surface

[0116] The predicted molecular weight of murine TANGO 202 proteinwithout modification and prior to cleavage of the signal sequence isabout 51.5 kilodaltons. The predicted molecular weight of the maturemurine TANGO 202 protein without modification and after cleavage of thesignal sequence is about 49.7 kilodaltons.

[0117] Human and murine TANGO 202 proteins exhibit considerable sequencesimilarity, as indicated herein in FIGS. 1J and 1K. FIGS. 1J and 1Kdepict an aligmnent of human and murine TANGO 202 amino acid sequences(SEQ ID NOs: 3 and 69, respectively). In this alignment (made using theALIGN software {Myers and Miller (1989) CABIOS, ver. 2.0}; pam120.matscoring matrix; gap penalties −12/−4), the proteins are 76.5% identical.The human and murine ORFs encoding TANGO 202 are 87.4% identical, asassessed using the same software and parameters.

[0118] In situ hybridization experiments in mouse tissues indicated thatmRNA corresponding to the cDNA encoding TANGO 202 is expressed in thetissues listed in Table III, wherein “+” indicates detectable expressionand “++” indicates a greater level of expression than “+”. TABLE IIIRelative Level of Animal Tissue Expression Mouse bladder, especially in++ (Adult) transitional epithelium renal glomeruli + brain + heart +liver + spleen + placenta + Mouse ubiquitous + (Embryo)

[0119] Biological Function of TANGO 202 Proteins, Nucleic Acids, andModulators Thereof

[0120] TANGO 202 proteins are involved in disorders which affect bothtissues in which they are normally expressed and tissues in which theyare normally not expressed. Based on the observation that TANGO 202 isexpressed in human fetal skin, ubiquitously in fetal mouse tissues, inadult murine bone marrow stromal cells, and in cells of adult murinebladder, renal glomeruli, brain, heart, liver, spleen and placenta,TANGO 202 protein is involved in one or more biological processes whichoccur in these tissues. In particular, TANGO 202 is involved inmodulating growth, proliferation, survival, differentiation, andactivity of cells of these tissues including, but not limited to,hematopoietic and fetal cells. Thus, TANGO 202 has a role in disorderswhich affect these cells and their growth, proliferation, survival,differentiation, and activity. Ubiquitous expression of TANGO 202 infetal murine tissues, contrasted with limited expression in adult murinetissues further indicates that TANGO 202 is involved in disorders inwhich it is inappropriately expressed (e.g., disorders in which TANGO202 is expressed in adult murine tissues other than bone marrow stromalcells and disorders in which TANGO 202 is not expressed in one or moredeveloping fetal tissues).

[0121] The presence of a Kringle domain in both the murine and humanTANGO 202 proteins indicates that this protein is involved in modulatingcellular binding to one or more mediators (e.g., proteins,phospholipids, intracellular organelles, or other cells), in modulatingproteolytic activity, or both. The presence of a Kringle domain in otherproteins (e.g., growth factors) indicates activities that these proteinsshare with TANGO 202 protein (e.g., modulating cell dissociation andmigration into and through extracellular matrices). The presence ofKringle domains in numerous plasma proteins, particularly coupled withthe observation that TANGO 202 is expressed in adult murine bone marrowstromal cells, indicates a role for TANGO 202 protein in modulatingbinding of blood or hematopoietic cells (or both) to one or moremediators. Thus, TANGO 202 is involved in disorders relating to aberrantcellular protease activity, inappropriate interaction or non-interactionof cells with mediators, and in blood and hematopoietic cell-relateddisorders. Such disorders include, by way of example and not limitation,immune disorders, infectious diseases, auto-immune disorders, vascularand cardiovascular disorders, disorders related to mal-expression ofgrowth factors, cancers, hematological disorders, and the like.

[0122] The cDNA encoding TANGO 202 exhibits significant nucleotidesequence similarity with a polynucleotide encoding akringle-domain-containing protein (designated HTHBZ47) described in theEuropean Patent Application No. EP 0 911 399 A2 (published Apr. 28,1999). Thus, the TANGO 202 protein can exhibit one or more of theactivities exhibited by HTHBZ47, and can be used to prevent, inhibit,diagnose, and treat one or more disorders for which HTHBZ47 is useful.These disorders include cancer, inflammation, autoimmune disorders,allergic disorders, asthma, rheumatoid arthritis, inflammation ofcentral nervous system tissues, cerebellar degeneration, Alzheimer'sdisease, Parkinson's disease, multiple sclerosis, amylotrophic lateralsclerosis, head injury damage and other neurological abnormalities,septic shock, sepsis, stroke, osteoporosis, osteoarthritis, ischemicreperfusion injury, cardiovascular disease, kidney disease, liverdisease, ischemic injury, myocardial infarction, hypotension,hypertension, AIDS, myelodysplastic syndromes and other hematologicabnormalities, aplastic anemia, male pattern baldness, and bacterial,fungal, protozoan, and viral infections.

[0123] The presence of a CUB domain in both the murine and human TANGO202 proteins indicates that this protein is involved in biologicalprocesses common to other CUB domain-containing proteins, such asdevelopmental processes and binding to mediators. Therefore, TANGO 202protein has a role in disorders which involve inappropriatedevelopmental processes (e.g., abnormally high proliferation orun-differentiation of a differentiated tissue or abnormally lowdifferentiation or proliferation of a non-developed ornon-differentiated tissue) and modulation of cell growth, proliferation,survival, differentiation, and activity. Such disorders include, by wayof example and not limitation, various cancers and birth anddevelopmental defects.

[0124] Thus, proteins and nucleic acids of the invention which areidentical to, similar to, or derived from human and murine TANGO 202proteins and nucleic acids encoding them are useful for preventing,diagnosing, and treating, among others, vascular and cardiovasculardisorders, hematological disorders, disorders related to mal-expressionof growth factors, and cancer. Other uses for these proteins and nucleicacids of the invention relate to modulating cell growth (e.g.,angiogenesis), proliferation (e.g., cancers), survival (e.g.,apoptosis), differentiation (e.g., hematopoiesis), and activity (e.g.,ligand-binding capacity). TANGO 202 proteins and nucleic acids encodingthem are also useful for modulating cell dissociation and modulatingmigration of cells in extracellular matrices.

[0125] TANGO 234

[0126] A cDNA clone (designated jthsa104d11) encoding at least a portionof human TANGO 234 protein was isolated from a human fetal spleen cDNAlibrary. The human TANGO 234 protein is predicted by structural analysisto be a transmembrane protein, although it can exist in a secreted formas well.

[0127] The full length of the cDNA encoding human TANGO 234 protein(FIG. 2; SEQ ID NO: 9) is 4628 nucleotide residues. The ORF of thiscDNA, nucleotide residues 28 to 4386 of SEQ ID NO: 9 (i.e., SEQ ID NO:10), encodes a 1453-amino acid transmembrane protein (FIG. 2; SEQ ID NO:11).

[0128] The invention thus includes purified human TANGO 234 protein,both in the form of the immature 1453 amino acid residue protein (SEQ IDNO: 11) and in the form of the mature 1413 amino acid residue protein(SEQ ID NO: 13). Mature human TANGO 234 protein can be synthesizedwithout the signal sequence polypeptide at the amino terminus thereof,or it can be synthesized by generating immature TANGO 234 protein andcleaving the signal sequence therefrom.

[0129] In addition to full length mature and immature human TANGO 234proteins, the invention includes fragments, derivatives, and variants ofthese TANGO 234 proteins, as described herein. These proteins,fragments, derivatives, and variants are collectively referred to hereinas polypeptides of the invention or proteins of the invention.

[0130] The invention also includes nucleic acid molecules which encode apolypeptide of the invention. Such nucleic acids include, for example, aDNA molecule having the nucleotide sequence listed in SEQ ID NO: 9 orsome portion thereof, such as the portion which encodes mature TANGO 234protein, immature TANGO 234 protein, or a domain of TANGO 234 protein.These nucleic acids are collectively referred to as nucleic acids of theinvention.

[0131] TANGO 234 proteins and nucleic acid molecules encoding themcomprise a family of molecules having certain conserved structural andfunctional features, as indicated by the conservation of amino acidsequence between human TANGO 234 protein and bovine WC1 protein, asshown in FIGS. 2K through 2P, and the conservation of nucleotidesequence between the ORFs encoding human TANGO 234 protein and bovineWC1 protein, as shown in FIGS. 2Q-1 through 2Q-17.

[0132] A common domain present in TANGO 234 proteins is a signalsequence. As used herein, a signal sequence includes a peptide of atleast about 10 amino acid residues in length which occurs at the aminoterminus of membrane-bound proteins and which contains at least about45% hydrophobic amino acid residues such as alanine, leucine,isoleucine, phenylalanine, proline, tyrosine, tryptophan, or valine. Ina preferred embodiment, a signal sequence contains at least about 10 to35 amino acid residues, preferably about 10 to 20 amino acid residues,and has at least about 35-60%, more preferably 40-50%, and morepreferably at least about 45% hydrophobic residues. A signal sequenceserves to direct a protein containing such a sequence to a lipidbilayer. Thus, in one embodiment, a TANGO 234 protein contains a signalsequence corresponding to amino acid residues 1 to 40 of SEQ ID NO: 11(SEQ ID NO: 12). The signal sequence is cleaved during processing of themature protein.

[0133] TANGO 234 proteins can include an extracellular domain. The humanTANGO 234 protein extracellular domain is located from about amino acidresidue 41 to about amino acid residue 1359 of SEQ ID NO: 3. TANGO 234can alternately exist in a secreted form, such as a mature proteinhaving the amino acid sequence of amino acid residues 41 to 1453 orresidues 41 to about 1359 of SEQ ID NO: 11.

[0134] In addition, TANGO 234 include a transmembrane domain. In oneembodiment, a TANGO 234 protein of the invention contains atransmembrane domain corresponding to about amino acid residues 1360 to1383 of SEQ ID NO: 11 (SEQ ID NO: 15).

[0135] The present invention includes TANGO 234 proteins having acytoplasmic domain, particularly including proteins having acarboxyl-terminal cytoplasmic domain. The human TANGO 234 cytoplasmicdomain is located from about amino acid residue 1384 to amino acidresidue 1453 of SEQ ID NO: 11 (SEQ ID NO: 16).

[0136] TANGO 234 proteins typically comprise a variety of potentialpost-translational modification sites (often within an extracellulardomain), such as those described herein in Table IV, as predicted bycomputerized sequence analysis of TANGO 234 proteins using amino acidsequence comparison software (comparing the amino acid sequence of TANGO234 with the information in the PROSITE database {rel. 12.2; February,1995} and the Hidden Markov Models database {Rel. PFAM 3.3 }). Incertain embodiments, a protein of the invention has at least 1, 2, 4, 6,10, 15, or 20 or more of the post-translational modification siteslisted in Table IV. TABLE IV Amino Acid Type of Potential ModificationSite or Residues of Amino Acid Domain SEQ ID NO: 11 SequenceN-glycosylation site 42 to 45 NGTD 78 to 81 NTTA 120 to 123 NESA 161 to164 NNSC 334 to 337 NESF 377 to 380 NCSG 441 to 444 NESA 548 to 551 NESN637 to 640 NAST 972 to 975 NESL 1013 to 1016 NVSD 1084 to 1087 NATV 11O4to 1107 NCTG 1161 to 1164 NGTW 1171 to 1174 NITT 1318 to 1321 NESF 1354to 1357 NASS Glycosaminoglycan attachment site 558 to 561 SGWG 665 to668 SGWG cAMP- or cGMP-dependent protein 1229 to 1232 RRIS kinasephosphorylation site 1399 to 1402 RRGS Protein kinase C phosphorylationsite 165 to 167 SGR 268 to 270 TNR 379 to 381 SGR 419 to 421 SRR 469 to471 SDK 506 to 508 STR 589 to 591 SNR 593 to 595 SGR 661 to 663 SCR 696to 698 SSR 746 to 748 TER 805 to 807 SGR 815 to 817 TWR 959 to 961 SVR1256 to 1258 SGR 1349 to 1351 SLK 1396 to 1398 STR Casein kinase IIphosphorylation site 44 to 47 TDLE 71 to 74 TVCD 178 to 181 TICD 245 to248 SHNE 253 to 256 TCYD 258 to 261 SDLE 319 to 322 SGSD 332 to 335 SGNE392 to 395 TICD 439 to 442 TGNE 606 to 609 TVCD 622 to 625 SQLD 673 to676 SHSE 686 to 689 SDME 760 to 763 TGGE 765 to 768 SLWD 818 to 821 SVCD845 to 848 SVGD 857 to 860 TWAE 907 to 910 SQCD 923 to 926 SLCD 927 to930 THWD 974 to 977 SLLD 1059 to 1062 TICD 1106 to 1109 TGTE 1145 to1148 SETE 1233 to 1236 SPAE 1241 to 1244 TCED 1269 to 1272 TVCD 1402 to1405 SLEE 1425 to 1428 TSDD N-myristoylation site 67 to 72 GQWGTV 90 to95 GCPFSF 101 to 106 GQAVTR 119 to 124 GNESAL 133 to 138 GSHNCY 160 to165 GNNSCS 197 to 202 GCPSSF 226 to 231 GNELAL 240 to 245 GNHDCS 267 to272 GTNRCM 304 to 309 GCGTAL 328 to 333 GVSCSG 374 to 379 GSNNCS 411 to416 GCPFSV 418 to 423 GSRRAK 440 to 445 GNESAL 465 to 470 GVICSD 547 to552 GNESNI 588 to 593 GSNRCS 632 to 637 GMGLGN 668 to 673 GNNDCS 679 to684 GVICSD 695 to 700 GSSRCA 712 to 717 GILCAN 720 to 725 GMNIAE 758 to763 GCTGGE 853 to 858 GNGLTW 891 to 896 GVVCSR 944 to 949 GTALST 985 to990 GAPPCI 992 to 997 GNTVSV 1078 to 1083 GCGVAF 1121 to 1126 GQHDCR1132 to 1137 GVICSE 1162 to 1167 GTWGSV 1185 to 1190 GCGENG 1265 to 1270GSWGTV 1288 to 1293 GCGSAL 1302 to 1307 GQGTGT 1331 to 1336 GQSDCG 1342to 1347 GVRCSG 1422 to 1427 GTRTSD 1443 to 1438 GCEDAS 1444 to 1449GVLPAS Amidation site 1167 to 1170 VGRR Speract receptor repeated (SRR)53 to 90 See FIG. 2 domain signature 160 to 197 See FIG. 2 267 to 304See FIG. 2 1041 to 1078 See FIG. 2 1251 to 1288 See FIG. 2 Scavengerreceptor cysteine-rich  51 to 148 See FIG. 2 (SRCR) domain 158 to 255See FIG. 2 265 to 362 See FIG. 2 372 to 469 See FIG. 2 479 to 576 SeeFIG. 2 586 to 683 See FIG. 2 693 to 790 See FIG. 2 798 to 895 See FIG. 2 903 to 1000 See FIG. 2 1039 to 1136 See FIG. 2 1146 to 1243 See FIG. 21249 to 1346 See FIG. 2

[0137] Among the domains that occur in TANGO 234 protein are SRR domainsand SRCR domains. In one embodiment, the protein of the invention has atleast one domain that is at least 55%, preferably at least about 65%,more preferably at least about 75%, yet more preferably at least about85%, and most preferably at least about 95% identical to one of thesedomains. In other embodiments, the protein has at least two of the SRRand SRCR domains described herein in Table IV. In other embodiments, theprotein has at least one SRR domain and at least one SRCR domain.

[0138] The SRR domain is named after a receptor domain identified in asea urchin egg protein designated speract. The consensus sequence ofthis domain (using standard one-letter amino acid codes, wherein X isany amino acid residue) is as follows.

-G-X₅-G-X₂-E-X₆-W-G-X₂-C-X₃-(F or Y or W)-X₈-C-X₃-G-.

[0139] Speract is a transmembrane glycoprotein of 500 amino acidresidues (Dangott et al. (1989) Proc. Natl. Acad. Sci. USA86:2128-2132). Structurally, this receptor consists of a largeextracellular domain of 450 residues, followed by a transmembrane regionand a small cytoplasmic domain of 12 amino acid residues. Theextracellular domain contains four repeats of an approximately 115 aminoacid domain. There are 17 amino acid residues that are perfectlyconserved in the four repeats in speract, including six cysteineresidues, six glycine residues, and two glutamate residues. TANGO 234has five SRR domains, in which 16 of the 17 conserved speract residuesare present of four of the SRR domains and 15 are present in theremaining SRR domain. This domain is designated the speract receptorrepeated domain. The amino acid sequence of mammalian macrophagescavenger receptor type I (MSRI) exhibits such a domain (Freeman et al.(1990) Proc. Natl. Acad. Sci. USA 87:8810-8814). MSRI proteins aremembrane glycoproteins implicated in the pathologic deposition ofcholesterol in arterial walls during atherogenesis. TANGO 234 isinvolved in one or more physiological processes related to cholesteroldeposition and atherogenesis, as well as other vascular andcardiovascular disorders.

[0140] Scavenger receptor cysteine-rich (SRCR) domains are disulfiderich extracellular domains which are present in certain cell surface andsecreted proteins. Proteins having SRCR domains exhibit diverse ligandbinding specificity. For example, in addition to modified lipoproteins,some of these proteins bind a variety of surface components ofpathogenic microorganisms, and some of the proteins bind apoptoticcells. SRCR domains are also involved in mediating immune developmentand response. Other SRCR-containing proteins are involved in binding ofmodified lipoproteins (e.g., oxidized low density lipoprotein {LDL}) byspecialized macrophages, leading to the formation of macrophages filledwith cholesteryl ester droplets (i.e., foam cells). TANGO 234 isinvolved in one or more physiological processes in which these otherSRCR domain-containing proteins are involved, such as LDL uptake andmetabolism, regulation of serum cholesterol level, atherogenesis,atherosclerosis, bacterial or viral infections, immune development, andgeneration and perseverance of immune responses.

[0141] WC1 is a ruminant protein having an SRCR domain. WC1 and gammadelta T-cell receptor are the only known gamma delta T-cell specificantigens. Antibodies which bind specifically with WC1 induce growtharrest in IL-2-dependent gamma delta T-cell and augment proliferation ofgamma delta T-cells in an autologous mixed lymphocyte reaction or in thepresence of anti-CD2 or anti-CD5 antibodies. Injection of antibodieswhich bind specifically with WC1 into calves results in long-lastingdepletion of gamma delta T-cells. Furthermore, antibodies which bindspecifically with WC1 can be used to purify gamma delta T-cells.

[0142] Gamma delta T-cells are involved in a variety of physiologicalprocesses. For example, these cells are potential mediators of allergicairway inflammation and lyme disease. Furthermore, these cells areinvolved in natural resistance to viral infections and can mediateautoimmune diseases. Elimination of gamma delta T-cells by injection ofantibodies which bind specifically therewith can affect the outcomes ofthese disorders.

[0143] TANGO 234 is likely the human orthologue of ruminant protein WC1,and thus is involved with the physiological processes described above inhumans. An alignment of the amino acid sequences of (human) TANGO 234and bovine WC1 protein is shown in FIGS. 2K-2P. In this alignment (madeusing the ALIGN software {Myers and Miller (1989) CABIOS, ver. 2.0};pam120.mat scoring matrix; gap penalties −12/−4), the proteins are 40.4%identical. An alignment of the nucleotide sequences of the ORFs encoding(human) TANGO 234 and bovine WC1 protein is shown in FIGS. 2Q-1 to2Q-17. The two ORFs are 54.3% identical, as assessed using the samesoftware and parameters.

[0144] The signal peptide prediction program SIGNALP (Nielsen et al.(1997) Protein Engineering 10:1-6) predicted that human TANGO 234protein includes a 40 amino acid signal peptide (amino acid residues 1to 40 of SEQ ID NO: 11; SEQ ID NO: 12) preceding the mature TANGO 234protein (amino acid residues 41 to 4386 of SEQ ID NO: 11; SEQ ID NO:13). Human TANGO 234 protein includes an extracellular domain (aminoacid residues 41 to 1359 of SEQ ID NO: 11; SEQ ID NO: 14); atransmembrane domain (amino acid residues 1360 to 1383 of SEQ ID NO: 11;SEQ ID NO: 15); and a cytoplasmic domain (amino acid residues 1384 to1453 of SEQ ID NO: 11; SEQ ID NO: 16).

[0145]FIG. 2J depicts a hydrophilicity plot of human TANGO 234 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Thehydrophobic region which corresponds to amino acid residues 1 to 40 ofSEQ ID NO: 11 is the signal sequence of human TANGO 234 (SEQ ID NO: 12).The hydrophobic region which corresponds to amino acid residues 1360 to1383 of SEQ ID NO: 11 is the transmembrane domain of human TANGO 234(SEQ ID NO: 15). As described elsewhere herein, relatively hydrophilicregions are generally located at or near the surface of a protein, andare more frequently effective immunogenic epitopes than are relativelyhydrophobic regions. For example, the region of human TANGO 234 proteinfrom about amino acid residue 225 to about amino acid residue 250appears to be located at or near the surface of the protein, while theregion from about amino acid residue 990 to about amino acid residue1000 appears not to be located at or near the surface.

[0146] The predicted molecular weight of human TANGO 234 protein withoutmodification and prior to cleavage of the signal sequence is about 159.3kilodaltons. The predicted molecular weight of the mature human TANGO234 protein without modification and after cleavage of the signalsequence is about 154.7 kilodaltons.

[0147] Chromosomal mapping to identify the location of the gene encodinghuman TANGO 234 protein indicated that the gene was located atchromosomal location h12p13 (with synteny to mo6). Flanking chromosomalmarkers include WI-6980 and GATA8A09.43. Nearby human loci include IBD2(inflammatory bowel disease 2), FPF (familial periodic fever), and HPDR2(hypophosphatemia vitamin D resistant rickets 2). Nearby genes are KLRC(killer cell receptor cluster), DRPLA (dentatorubro-pallidoluysianatrophy), GAPD (glyceraldehyde-3-phosphate) dehydrogenase, and PXR1(peroxisome receptor 1). Murine chromosomal mapping indicated that themurine orthologue is located near the scr (scruffy) locus. Nearby mousegenes include drpla (dentatorubral phillidoluysian atrophy), prp(proline rich protein), and kap (kidney androgen regulated protein).

[0148] Northern analysis experiments indicated that mRNA correspondingto the cDNA encoding TANGO 234 is expressed in the tissues listed inTable V, wherein “++” indicates moderate expression, “+” indicates lowerexpression, and “−” indicates no detectable expression. TABLE V AnimalTissue Relative Level of Expression Human spleen ++ fetal lung ++ lung +thymus + bone marrow − peripheral blood leukocytes −

[0149] Biological Function of TANGO 234 Proteins, Nucleic Acids, andModulators Thereof

[0150] TANGO 234 proteins are involved in disorders which affect bothtissues in which they are normally expressed and tissues in which theyare normally not expressed. Based on the observation that TANGO 234 isexpressed in human fetal lung, spleen, and, to a lesser extent in adultlung and thymus tissue, TANGO 234 protein is involved in one or morebiological processes which occur in these tissues. In particular, TANGO234 is involved in modulating growth, proliferation, survival,differentiation, and activity of cells including, but not limited to,lung, spleen, thymus bone marrow, hematopoietic, peripheral bloodleukocytes, and fetal cells of the animal in which it is normallyexpressed. Thus, TANGO 234 has a role in disorders which affect thesecells and their growth, proliferation, survival, differentiation, andactivity. Expression of TANGO 234 in an animal is also involved inmodulating growth, proliferation, survival, differentiation, andactivity of cells and viruses which are foreign to the host (i.e.,bacterial, fungal, and viral infections).

[0151] Homology of human TANGO 234 with bovine WC1 protein indicatesthat TANGO 234 has physiological functions in humans analogous to thefunctions of WC1 in ruminants. Thus, TANGO 234 is involved in modulatinggrowth, proliferation, survival, differentiation, and activity of gammadelta T cells. For example, TANGO 234 affects the ability of gamma deltaT cells to interact with chemokines such as interleukin-2. TANGO 234therefore is involved in the physiological processes associated withallergic airway inflammation, lyme arthritis, resistance to viralinfection, auto-immune diseases, and the like.

[0152] In addition, presence in TANGO 234 of SRR and SRCR domainsindicates that TANGO 234 is involved in physiological functionsidentical or analogous to the functions performed by other proteinshaving such domains. For example, like other SRR domain-containingproteins, TANGO 234 modulates cholesterol deposition in arterial walls,and is thus involved in development and persistence of atherogenesis andarteriosclerosis, as well as other vascular and cardiovasculardisorders. Like other SRCR domain-containing proteins, TANGO 234 isinvolved in uptake and metabolism of LDL, regulation of serumcholesterol level, and can modulate these processes as well as theprocesses of atherogenesis, arteriosclerosis, immune development, andgeneration and perseverance of immune responses to bacterial, fungal,and viral infections.

[0153] TANGO 265

[0154] A cDNA clone (designated jthsa079g01) encoding at least a portionof human TANGO 265 protein was isolated from a human fetal spleen cDNAlibrary. The human TANGO 265 protein is predicted by structural analysisto be a transmembrane membrane protein, although it can exist in asecreted form as well.

[0155] The full length of the cDNA encoding human TANGO 265 protein(FIG. 3; SEQ ID NO: 17) is 3104 nucleotide residues. The ORF of thiscDNA, nucleotide residues 32 to 2314 of SEQ ID NO: 17 (i.e., SEQ ID NO:18), encodes a 761-amino acid transmembrane protein (FIG. 3; SEQ ID NO:19).

[0156] The invention thus includes purified TANGO 265 protein, both inthe form of the immature 761 amino acid residue protein (SEQ ID NO: 19)and in the form of the mature 730 amino acid residue protein (SEQ ID NO:21). Mature TANGO 265 protein can be synthesized without the signalsequence polypeptide at the amino terminus thereof, or it can besynthesized by generating immature TANGO 265 protein and cleaving thesignal sequence therefrom.

[0157] In addition to full length mature and immature TANGO 265proteins, the invention includes fragments, derivatives, and variants ofTANGO 265 protein, as described herein. These proteins, fragments,derivatives, and variants are collectively referred to herein aspolypeptides of the invention or proteins of the invention.

[0158] The invention also includes nucleic acid molecules which encode apolypeptide of the invention. Such nucleic acids include, for example, aDNA molecule having the nucleotide sequence listed in SEQ ID NO: 17 orsome portion thereof, such as the portion which encodes mature TANGO 265protein, immature TANGO 265 protein, or a domain of TANGO 265 protein.These nucleic acids are collectively referred to as nucleic acids of theinvention.

[0159] TANGO 265 proteins and nucleic acid molecules encoding themcomprise a family of molecules having certain conserved structural andfunctional features.

[0160] A common domain present in TANGO 265 proteins is a signalsequence. As used herein, a signal sequence includes a peptide of atleast about 10 amino acid residues in length which occurs at the aminoterminus of membrane-bound proteins and which contains at least about45% hydrophobic amino acid residues such as alanine, leucine,isoleucine, phenylalanine, proline, tyrosine, tryptophan, or valine. Ina preferred embodiment, a signal sequence contains at least about 10 to35 amino acid residues, preferably about 10 to 20 amino acid residues,and has at least about 35-60%, more preferably 40-50%, and morepreferably at least about 45% hydrophobic residues. A signal sequenceserves to direct a protein containing such a sequence to a lipidbilayer. Thus, in one embodiment, a TANGO 265 protein contains a signalsequence corresponding to amino acid residues 1 to 31 of SEQ ID NO: 19(SEQ ID NO: 20). The signal sequence is cleaved during processing of themature protein

[0161] TANGO 265 proteins can also include an extracellular domain. Thehuman TANGO 265 protein extracellular domain is located from about aminoacid residue 32 to about amino acid residue 683 of SEQ ID NO: 17. TANGO265 can alternately exist in a secreted form, such as a mature proteinhaving the amino acid sequence of amino acid residues 32 to 761 orresidues 32 to about 683 of SEQ ID NO: 19.

[0162] TANGO 265 proteins can also include a transmembrane domain. Inone embodiment, a TANGO 265 protein of the invention contains atransmembrane domain corresponding to about amino acid residues 684 to704 of SEQ ID NO: 19 (SEQ ID NO: 23).

[0163] In addition, TANGO 265 proteins include a cytoplasmic domain,particularly including proteins having a carboxyl-terminal cytoplasmicdomain. The human TANGO 265 cytoplasmic domain is located from aboutamino acid residue 705 to amino acid residue 761 of SEQ ID NO: 19 (SEQID NO: 24).

[0164] TANGO 265 proteins typically comprise a variety of potentialpost-translational modification sites (often within an extracellulardomain), such as those described herein in Table VI, as predicted bycomputerized sequence analysis of TANGO 265 proteins using amino acidsequence comparison software (comparing the amino acid sequence of TANGO265 with the information in the PROSITE database {rel. 12.2; February,1995} and the Hidden Markov Models database {Rel. PFAM 3.3 }). Incertain embodiments, a protein of the invention has at least 1, 2, 4, 6,10, 15, or 20 or more of the post-translational modification siteslisted in Table VI. TABLE VI Amino Acid Type of Potential ModificationSite or Residues of Amino Acid Domain SEQ ID NO: 19 SequenceN-glycosylation site 120 to 123 NETQ 135 to 138 NVTH 496 to 499 NCSV 607to 610 NGLS Glycosaminoglycan attachment site 70 to 73 SGDG cAMP- orcGMP-dependent protein 108 to 111 RKKS kinase phosphorylation site 116to 119 KKKS 281 to 284 KKWT Protein kinase C phosphorylation site 106 to108 SDR 262 to 264 TSR 361 to 363 TSR 366 to 368 TYR 385 to 387 SDK 533to 535 SWK 555 to 557 SLR 721 to 723 TLR 738 to 740 SPK Casein kinase IIphosphorylation site 152 to 155 TFIE 176 to 179 SPFD 250 to 253 TASE 342to 345 SLLD 411 to 414 SGVE 498 to 501 SVYE 502 to 505 SCVD 574 to 577SILE 738 to 741 SPKE 745 to 748 SASD N-myristoylation site 79 to 84GAREAI 191 to 196 GMLYSG 331 to 336 GGTRSS 412 to 417 GVEYTR 437 to 442GTTTGS 620 to 625 GLYQCW 671 to 676 GAALAA Sema domain  64 to 478 SeeFIG. 3

[0165] An exemplary domains which occurs in TANGO 265 proteins is a semadomain. In one embodiment, the protein of the invention has at least onedomain that is at least 55%, preferably at least about 65%, morepreferably at least about 75%, yet more preferably at least about 85%,and most preferably at least about 95% identical to one of the semadomains described herein in Table VI.

[0166] Sema domains occur in semaphorin proteins. Semaphorins are alarge family of secreted and transmembrane proteins, some of whichfunction as repellent signals during neural axon guidance. The semadomain and a variety of semaphorin proteins in which it occurs aredescribed, for example, in Winberg et al. (1998 Cell 95:903-916). Semadomains also occur in human hepatocyte growth factor receptor (SwissprotAccession no. P08581) and the similar neuronal and epithelialtransmembrane receptor protein (Swissprot Accession no. P51805). Thepresence of an sema domain in human TANGO 265 protein indicates thatTANGO 265 is involved in one or more physiological processes in whichthe semaphorins are involved, has biological activity in common with oneor more of the semaphorins, or both.

[0167] Human TANGO 265 protein exhibits considerable sequence similarityto murine semaphorin B protein (GenBank Accession no. X85991), asindicated herein in FIGS. 3F-3H. FIGS. 3F-3H depict an alignment of theamino acid sequences of human TANGO 265 protein (SEQ ID NO: 19) andmurine semaphorin B protein (SEQ ID NO: 76). In this alignment(pam120.mat scoring matrix, gap penalties −12/−4), the amino acidsequences of the proteins are 82.3% identical. FIGS. 31 through 3Tdepict an alignment of the nucleotide sequences of cDNA encoding humanTANGO 265 protein (SEQ ID NO: 17) and murine cDNA encoding semaphorin Bprotein (SEQ ID NO: 77). In this alignment (Pam120.mat scoring matrix,gap penalties −12/−4), the nucleic acid sequences of the cDNAs are 76.2%identical. Thus, TANGO 265 is the human orthologue of murine semaphorinB and shares functional similarities to that protein.

[0168] It is known that semaphorins are bi-functional, capable offunctioning either as attractive axonal guidance proteins or asrepellent axonal guidance proteins (Wong et al. (1997) Development124:3597-3607). Furthermore, semaphorins bind with neuronal cell surfaceproteins designated plexins, which are expressed on both neuronal cellsand cells of the immune system (Comeau et al. (1998) Immunity 8:473-482;Jin and Strittmatter (1997) J. Neurosci. 17:6256-6263).

[0169] The signal peptide prediction program SIGNALP (Nielsen et al.(1997) Protein Engineering 10:1-6) predicted that human TANGO 265protein includes a 31 amino acid signal peptide (amino acid residues 1to 31 of SEQ ID NO: 19; SEQ ID NO: 20) preceding the mature TANGO 265protein (amino acid residues 32 to 761 of SEQ ID NO: 19; SEQ ID NO: 21).Human TANGO 265 protein includes an extracellular domain (amino acidresidues 32 to 683 of SEQ ID NO: 19; SEQ ID NO: 22); a transmembranedomain (amino acid residues 684 to 704 of SEQ ID NO: 19; SEQ ID NO: 23);and a cytoplasmic domain (amino acid residues 705 to 761 of SEQ ID NO:19; SEQ ID NO: 24).

[0170]FIG. 3U depicts a hydrophilicity plot of human TANGO 265 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Thehydrophobic region which corresponds to amino acid residues 1 to 31 ofSEQ ID NO: 19 is the signal sequence of human TANGO 265 (SEQ ID NO: 20).The hydrophobic region which corresponds to amino acid residues 684 to704 of SEQ ID NO: 19 is the transmembrane domain of human TANGO 265 (SEQID NO: 23). As described elsewhere herein, relatively hydrophilicregions are generally located at or near the surface of a protein, andare more frequently effective immunogenic epitopes than are relativelyhydrophobic regions. For example, the region of human TANGO 265 proteinfrom about amino acid residue 350 to about amino acid residue 375appears to be located at or near the surface of the protein, while theregion from about amino acid residue 230 to about amino acid residue 250appears not to be located at or near the surface.

[0171] The predicted molecular weight of human TANGO 265 protein withoutmodification and prior to cleavage of the signal sequence is about 83.6kilodaltons. The predicted molecular weight of the mature human TANGO265 protein without modification and after cleavage of the signalsequence is about 80.2 kilodaltons.

[0172] Chromosomal mapping was performed by computerized comparison ofTANGO 265 cDNA sequences against a chromosomal mapping database in orderto identify the approximate location of the gene encoding human TANGO265 protein. This analysis indicated that the gene was located onchromosome 1 between markers D1S305 and D1S2635.

[0173] Biological Function of TANGO 265 Proteins, Nucleic Acids, andModulators Thereof

[0174] TANGO 265 proteins are involved in disorders which affect bothtissues in which they are normally expressed and tissues in which theyare normally not expressed. Based on the observation that TANGO 265 isexpressed in human fetal spleen, involvement of TANGO 202 protein inimmune system development and modulation is indicated.

[0175] The presence of the sema domain in TANGO 265 indicates that thisprotein is involved in development of neuronal and epithelial tissuesand also functions as a repellant protein which guides axonaldevelopment. TANGO 265 modulates nerve growth and regeneration and alsomodulates growth and regeneration of other epithelial tissues.

[0176] The observation that TANGO 265 shares significant identity withmurine semaphorin B suggests that it has activity identical or analogousto the activity of this protein. These observations indicate that TANGO265 modulates growth, proliferation, survival, differentiation, andactivity of neuronal cells and immune system cells. Thus, TANGO 265protein is useful, for example, for guiding neural axon development, formodulating differentiation of cells of the immune system, for modulatingcytokine production by cells of the immune system, for modulatingreactivity of cells of the immune system toward cytokines, formodulating initiation and persistence of an inflammatory response, andfor modulating proliferation of epithelial cells.

[0177] TANGO 273

[0178] A cDNA clone (designated jthoc028g06) encoding at least a portionof human TANGO 273 protein was isolated from alipopolysaccharide-(LPS-)stimulated human osteoblast cDNA library. Thecorresponding murine cDNA clone (designated jtmoa001c04) was isolatedfrom an LPS-stimulated murine osteoblast cDNA library. The human andmurine TANGO 273 proteins are predicted by structural analysis to betransmembrane proteins.

[0179] The full length of the cDNA encoding human TANGO 273 protein(FIG. 4; SEQ ID NO: 25) is 2964 nucleotide residues. The ORF of thiscDNA, nucleotide residues 135 to 650 of SEQ ID NO: 25 (i.e., SEQ ID NO:26), encodes a 172-amino acid transmembrane protein (FIG. 4; SEQ ID NO:27).

[0180] The invention thus includes purified human TANGO 273 protein,both in the form of the immature 172 amino acid residue protein (SEQ IDNO: 27) and in the form of the mature 150 amino acid residue protein(SEQ ID NO: 29). The invention also includes purified murine TANGO 273protein, both in the form of the immature 172 amino acid residue protein(SEQ ID NO: 74) and in the form of the mature 150 amino acid residueprotein (SEQ ID NO: 44). Mature human or murine TANGO 273 proteins canbe synthesized without the signal sequence polypeptide at the aminoterminus thereof, or they can be synthesized by generating immatureTANGO 273 protein and cleaving the signal sequence therefrom.

[0181] In addition to full length mature and immature human and murineTANGO 273 proteins, the invention includes fragments, derivatives, andvariants of these TANGO 273 proteins, as described herein. Theseproteins, fragments, derivatives, and variants are collectively referredto herein as polypeptides of the invention or proteins of the invention.

[0182] The invention also includes nucleic acid molecules which encode apolypeptide of the invention. Such nucleic acids include, for example, aDNA molecule having the nucleotide sequence listed in SEQ ID NO: 25 orsome portion thereof or SEQ ID NO: 73 or some portion thereof, such asthe portion which encodes mature TANGO 273 protein, immature TANGO 273protein, or a domain of TANGO 273 protein. These nucleic acids arecollectively referred to as nucleic acids of the invention.

[0183] TANGO 273 proteins and nucleic acid molecules encoding themcomprise a family of molecules having certain conserved structural andfunctional features. This family includes, by way of example, the humanand murine TANGO 273 proteins.

[0184] A common domain of TANGO 273 proteins is a signal sequence. Asused herein, a signal sequence includes a peptide of at least about 10amino acid residues in length which occurs at the amino terminus ofmembrane-bound proteins and which contains at least about 45%hydrophobic amino acid residues such as alanine, leucine, isoleucine,phenylalanine, proline, tyrosine, tryptophan, or valine. In a preferredembodiment, a signal sequence contains at least about 10 to 35 aminoacid residues, preferably about 10 to 20 amino acid residues, and has atleast about 35-60%, more preferably 40-50%, and more preferably at leastabout 45% hydrophobic residues. A signal sequence serves to direct aprotein containing such a sequence to a lipid bilayer. Thus, in oneembodiment, a TANGO 273 protein contains a signal sequence correspondingto amino acid residues 1 to 22 of SEQ ID NO: 27 (SEQ ID NO: 28) or toamino acid residues 1 to 22 of SEQ ID NO: 74. The signal sequence iscleaved during processing of the mature protein.

[0185] TANGO 273 proteins can also include an extracellular domain. Thehuman TANGO 273 protein extracellular domain is located from about aminoacid residue 23 to about amino acid residue 60 of SEQ ID NO: 27, and themurine TANGO 273 protein extracellular domain is located from aboutamino acid residue 23 to about amino acid residue 60 of SEQ ID NO: 74.

[0186] The present invention also includes TANGO 273 proteins having atransmembrane domain. As used herein, a “transmembrane domain” refers toan amino acid sequence having at least about 15 to 30 amino acidresidues in length and which contains at least about 65-70% hydrophobicamino acid residues such as alanine, leucine, phenylalanine, protein,tyrosine, tryptophan, or valine. In a preferred embodiment, atransmembrane domain contains at least about 15 to 20 amino acidresidues, preferably about 20 to 25 amino acid residues, and has atleast about 60-80%, more preferably 65-75%, and more preferably at leastabout 70% hydrophobic residues. Thus, in one embodiment, a human TANGO273 protein of the invention contains a transmembrane domaincorresponding to about amino acid residues 61 to 81 of SEQ ID NO: 27(SEQ ID NO: 31). In another embodiment, a murine TANGO 273 protein ofthe invention contains a transmembrane domain corresponding to aboutamino acid residues 61 to 81 of SEQ ID NO: 74.

[0187] In addition, TANGO 273 proteins include a cytoplasmic domain. Thehuman TANGO 273 cytoplasmic domain is located from about amino acidresidue 82 to amino acid residue 172 of SEQ ID NO: 27 (SEQ ID NO: 32),and the murine TANGO 273 cytoplasmic domain is located from about aminoacid residue 82 to amino acid residue 172 of SEQ ID NO: 74.

[0188] TANGO 273 proteins typically comprise a variety of potentialpost-translational modification sites (often within an extracellulardomain), such as those described herein in Tables VII and VIII, aspredicted by computerized sequence analysis of human and murine TANGO273 proteins using amino acid sequence comparison software (comparingthe amino acid sequence of TANGO 273 with the information in the PROSITEdatabase {rel. 12.2; February, 1995} and the Hidden Markov Modelsdatabase {Rel. PFAM 3.3}). In certain embodiments, a protein of theinvention has at least 1, 2, 3, 4, 5, or all 6 of the post-translationalmodification sites listed in Table VII. In other embodiments, theprotein of the invention has at least 1, 2, 3, 4, 5, 6, or all 7 of thepost-translational modification sites listed in Table VIII. TABLE VIIAmino Acid Type of Potential Modification Site or Residues of Amino AcidDomain SEQ ID NO: 27 Sequence N-glycosylation site  97 to 100 NVSYCasein kinase II phosphorylation site 41 to 44 SYED N-myristoylationsite 31 to 36 GLYPTY 47 to 52 GSRCCV 70 to 75 GVLFCC 131 to 136 GNSMAMSrc Homology 3 (SH3) domain binding 86 to 90 YPPPL site 103 to 107 QPPNP113 to 117 QPGPP 121 to 125 DPGGP 140 to 145 VPPNSP 151 to 155 CPPPP 160to 164 TPPPP

[0189] TABLE VIII Amino Acid Type of Potential Modification Site orResidues of Amino Acid Domain SEQ ID NO: 74 Sequence N-glycosylationsite 97 to 100 NVSY Casein kinase II phosphorylation site 41 to 44 SYEDN-myristoylation site 31 to 36 GLYPTY 47 to 52 GSRCCV 70 to 75 GVLFCC131 to 136 GNTMAM Src Homology 3 (SH3) domain binding 86 to 90 YPPPLsite 103 to 107 QPPNP 115 to 119 GPPYY 121 to 125 DPGGP 141 to 145 QPNSP151 to 155 YPPPP 160 to 164 TPPPP Amidation site 1 to 4 MGRR

[0190] The amino acid sequence of TANGO 273 protein includes about sevenpotential proline-rich Src homology 3 (SH3) domain binding sites nearerthe cytoplasmic portion of the protein. SH3 domains mediate specificassembly of protein complexes, presumably by interacting withproline-rich protein domains (Morton and Campbell (1994) Curr. Biol.4:615-617). SH3 domains also mediate interactions between proteinsinvolved in transmembrane signal transduction. Coupling of proteinsmediated by SH3 domains has been implicated in a variety ofphysiological systems, including those involving regulation of cellgrowth and proliferation, endocytosis, and activation of respiratoryburst.

[0191] SH3 domains have been described in the art (e.g., Mayer et al.(1988) Nature 332:272-275; Musacchio et al. (1992) FEBS Lett. 307:55-61;Pawson and Schlessinger (1993) Curr. Biol. 3:434-442; Mayer andBaltimore (1993) Trends Cell Biol. 3:8-13; Pawson (1993) Nature373:573-580), and occur in a variety of cytoplasmic proteins, includingseveral (e.g., protein tyrosine kinases) involved in transmembranesignal transduction. Among the proteins in which one or more SH3 domainsoccur are protein tyrosine kinases such as those of the Src, Abl, Bkt,Csk and ZAP70 families, mammalian phosphatidylinositol-specificphospholipases C-gamma-1 and -2, mammalian phosphatidylinositol 3-kinaseregulatory p85 subunit, mammalian Ras GTPase-activating protein (GAP),proteins which mediate binding of guanine nucleotide exchange factorsand growth factor receptors (e.g., vertebrate GRB2, Caenorhabditiselegans sem-5, and Drosophila DRK proteins), mammalian Vav oncoprotein,guanidine nucleotide releasing factors of the CDC 25 family (e.g., yeastCDC25, yeast SCD25, and fission yeast ste6 proteins), MAGUK proteins(e.g., mammalian tight junction protein ZO-1, vertebrate erythrocytemembrane protein p55, C. elegans protein lin-2, rat protein CASK, andmammalian synaptic proteins SAP90/PSD-95, CHAPSYN-110/PSD-93,SAP97/DLG1, and SAP102), proteins which interact with vertebratereceptor protein tyrosine kinases (e.g., mammalian cytoplasmic proteinNck and oncoprotein Crk), chicken Src substrate p80/85 protein(cortactin), human hemopoietic lineage cell specific protein Hs1,mammalian dihydrouridine-sensitive L-type calcium channel beta subunit,human myasthenic syndrome antigen B (MSYB), mammalian neutrophilcytosolic activators of NADPH oxidase (e.g., p47 {NCF-1}, p67 {NCF-2},and C. elegans protein B0303.7) myosin heavy chains (MYO3) from amoebae,from slime molds, and from yeast, vertebrate and Drosophila spectrin andfodrin alpha chain proteins, human amphiphysin, yeast actin-bindingproteins ABP1 and SLA3, yeast protein BEM 1, fission yeast protein scd2(ra13), yeast BEM1-binding proteins BOI2 (BEB1) and BOB1 (BOI1), yeastfusion protein FUS1, yeast protein RSV167, yeast protein SSU81, yeasthypothetical proteins YAR014c, YFR024c, YHL002w, YHR016c, YJL020C, andYHR114w, hypothetical fission yeast protein SpAC12C2.05c, and C. eleganshypothetical protein F42H10.3. Of these proteins, multiple SH3 domainsoccur in vertebrate GRB2 protein, C. elegans sem-5 protein, DrosophilaDRK protein, oncoprotein Crk, mammalian neutrophil cytosolic activatorsof NADPH oxidase p47 and p67, yeast protein BEM1, fission yeast proteinscd2, yeast hypothetical protein YHR114w, mammalian cytoplasmic proteinNck, C. elegans neutrophil cytosolic activator of NADPH oxidase B0303.7,and yeast actin-binding protein SLA1. Of these proteins, three or moreSH3 domains occur in mammalian cytoplasmic protein Nck, C. elegansneutrophil cytosolic activator of NADPH oxidase B0303.7, and yeastactin-binding protein SLA1. The presence of SH3 domain binding sites inTANGO 273 indicates that TANGO 273 interacts with one or more of theseand other SH3 domain-containing proteins and is thus involved inphysiological processes in which one or more of these or other SH3domain-containing proteins are involved.

[0192] The signal peptide prediction program SIGNALP (Nielsen et al.(1997) Protein Engineering 10:1-6) predicted that human TANGO 273protein includes a 22 amino acid signal peptide (amino acid residues 1to 22 of SEQ ID NO: 27; SEQ ID NO: 28) preceding the mature TANGO 273protein (amino acid residues 23 to 172 of SEQ ID NO: 27; SEQ ID NO: 29).Human TANGO 273 protein includes an extracellular domain (amino acidresidues 23 to 60 of SEQ ID NO: 27; SEQ ID NO: 30); a transmembranedomain (amino acid residues 61 to 81 of SEQ ID NO: 27; SEQ ID NO: 31);and a cytoplasmic domain (amino acid residues 82 to 172 of SEQ ID NO:27; SEQ ID NO: 32).

[0193]FIG. 4I depicts a hydrophilicity plot of human TANGO 273 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Thehydrophobic region which corresponds to amino acid residues 1 to 22 ofSEQ ID NO: 27 is the signal sequence of human TANGO 273 (SEQ ID NO: 28).The hydrophobic region which corresponds to amino acid residues 61 to 81of SEQ ID NO: 27 is the transmembrane domain of human TANGO 273 (SEQ IDNO: 31). As described elsewhere herein, relatively hydrophilic regionsare generally located at or near the surface of a protein, and are morefrequently effective immunogenic epitopes than are relativelyhydrophobic regions. For example, the region of human TANGO 273 proteinfrom about amino acid residue 100 to about amino acid residue 120appears to be located at or near the surface of the protein, while theregion from about amino acid residue 130 to about amino acid residue 140appears not to be located at or near the surface.

[0194] Chromosomal mapping was performed by computerized comparison ofTANGO 273 cDNA sequences against a chromosomal mapping database in orderto identify the approximate location of the gene encoding human TANGO273 protein. This analysis indicated that the gene was located onchromosome 7 between markers D7S2467 and D7S2552.

[0195] The predicted molecular weight of human TANGO 273 protein withoutmodification and prior to cleavage of the signal sequence is about 19.2kilodaltons. The predicted molecular weight of the mature human TANGO273 protein without modification and after cleavage of the signalsequence is about 16.8 kilodaltons.

[0196] Northern analysis experiments indicated that mRNA correspondingto the cDNA encoding TANGO 273 is expressed in the tissues listed inTable VIIa, wherein “++” indicates moderate expression and “+” indicateslower expression. TABLE VIIa Animal Tissue Relative Level of ExpressionHuman heart ++ brain ++ skeletal muscle ++ pancreas ++ placenta + lung +liver + kidney +

[0197] The full length of the cDNA encoding murine TANGO 273 protein(FIG. 4; SEQ ID NO: 72) is 2915 nucleotide residues. The ORF of thiscDNA, nucleotide residues 137 to 650 of SEQ ID NO: 72 (i.e., SEQ ID NO:73), encodes a 172-amino acid transmembrane protein (FIG. 4; SEQ ID NO:74).

[0198] The signal peptide prediction program SIGNALP (Nielsen et al.(1997) Protein Engineering 10:1-6) predicted that murine TANGO 273protein includes a 22 amino acid signal peptide (amino acid residues 1to 22 of SEQ ID NO: 74) preceding the mature TANGO 273 protein (aminoacid residues 23 to 172 of SEQ ID NO: 74; SEQ ID NO: 44). Murine TANGO273 protein includes an extracellular domain (amino acid residues 23 to60 of SEQ ID NO: 74); a transmembrane domain (amino acid residues 61 to81 of SEQ ID NO: 74); and a cytoplasmic domain (amino acid residues 82to 172 of SEQ ID NO: 74).

[0199]FIG. 4J depicts a hydrophilicity plot of murine TANGO 273 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Thehydrophobic region which corresponds to amino acid residues 1 to 22 ofSEQ ID NO: 74 is the signal sequence of murine TANGO 273. As describedelsewhere herein, relatively hydrophilic regions are generally locatedat or near the surface of a protein, and are more frequently effectiveimmunogenic epitopes than are relatively hydrophobic regions. Forexample, the region of murine TANGO 273 protein from about amino acidresidue 100 to about amino acid residue 120 appears to be located at ornear the surface of the protein, while the region from about amino acidresidue 130 to about amino acid residue 140 appears not to be located ator near the surface.

[0200] The predicted molecular weight of murine TANGO 273 proteinwithout modification and prior to cleavage of the signal sequence isabout 19.4 kilodaltons. The predicted molecular weight of the maturemurine TANGO 273 protein without modification and after cleavage of thesignal sequence is about 17.1 kilodaltons.

[0201] In situ analysis of murine TANGO 273 mRNA indicated that TANGO273 is expressed with central nervous system (CNS) tissues duringembryogenesis and into adulthood. Expression of TANGO 273 is widelyobserved in murine CNS tissues, including brain, spinal cord, eye, andolfactory epithelium at all embryonic ages examined (i.e., at embryonicdays 13.5, 14.5, 15.5, 16.5, and 18.5 and at post-natal day 1.5).

[0202] Human and murine TANGO 273 cDNA sequences exhibit significantnucleotide sequence identity with an expressed sequence tag (EST)isolated from a library of ESTs corresponding to proteins secreted fromprostate tissue, as described in PCT publication number WO 99/06550,published Feb. 11, 1999.

[0203] Human and murine TANGO 273 proteins exhibit considerable sequencesimilarity, as indicated herein in FIG. 4H. FIG. 4H depicts an alignmentof human and murine TANGO 273 protein amino acid sequences (SEQ ID NOs:27 and 74, respectively). In this alignment (pam120.mat scoring matrix,gap penalties −12/−4), the proteins are 89.5% identical. Alignment ofthe ORF encoding human TANGO 273 protein and the ORF encoding murineTANGO 273 protein using the same software and parameters indicated thatthe nucleotide sequences are 84.1% identical.

[0204] Biological Function of TANGO 273 Proteins, Nucleic Acids, andModulators Thereof

[0205] cDNAs encoding the human and murine TANGO 273 proteins were eachisolated from LPS-stimulated osteoblast cDNA libraries. These proteinsare involved in bone-related metabolism, homeostasis, and developmentdisorders. Thus, proteins and nucleic acids of the invention which areidentical to, similar to, or derived from human and murine TANGO 273proteins and nucleic acids encoding them are useful for preventing,diagnosing, and treating, among others, bone-related disorders such asosteoporosis, cancer, skeletal development disorders, bone fragility,and the like.

[0206] Expression of TANGO 273 in heart, brain, skeletal muscle, andpancreas, placenta, lung, liver, and kidney tissues is an indicationthat TANGO 273 proteins, nucleic acids encoding them, and agents thatmodulate activity or expression of either of these can be used tomodulate growth, proliferation, survival, differentiation, adhesion, andactivity of cells of these tissues, or to prognosticate, diagnose, andtreat one or more disorders which affect these tissues.

[0207] The fact that TANGO 273 is expressed at high levels inneurological tissues is an indication that TANGO 273 proteins, nucleicacids, and modulators thereof can be used to modulate proliferation,differentiation, or function of neurological cells in these tissues(e.g., neuronal cells). Thus, TANGO 273 proteins, nucleic acids, andmodulators thereof can be used to prognosticate, diagnose, and treat oneor more neurological disorders. Examples of such disorders include CNSdisorders, CNS-related disorders, focal brain disorders, global-diffusecerebral disorders, and other neurological and cerebrovasculardisorders.

[0208] CNS disorders include, but are not limited to cognitive andneurodegenerative disorders such as Alzheimer's disease, seniledementia, Huntington's disease, amyotrophic lateral sclerosis, andParkinson's disease, as well as Gilles de la Tourette's syndrome,autonomic function disorders such as hypertension and sleep disorders(e.g., insomnia, hypersomnia, parasomnia, and sleep apnea);neuropsychiatric disorders (e.g., schizophrenia, schizoaffectivedisorder, attention deficit disorder, dysthymic disorder, majordepressive disorder, mania, and obsessive-compulsive disorder);psychoactive substance use disorders; anxiety; panic disorder; andbipolar affective disorders (e.g., severe bipolar affective disorder andbipolar affective disorder with hypomania and major depression).

[0209] CNS-related disorders include disorders associated withdevelopmental, cognitive, and autonomic neural and neurologicalprocesses, such as pain, appetite, long term memory, and short termmemory.

[0210] Exemplary focal brain disorders include aphasia, apraxia,agnosia, and amnesias (e.g., posttraumatic amnesia, transient globalamnesia, and psychogenic amnesia). Global-diffuse cerebral disorderswith which TANGO 273 can be associated include coma, stupor,obtundation, and disorders of the reticular formation.

[0211] Other neurological disorders with which TANGO 273 can beassociated include ischemic syndromes (e.g., stroke), hypertensiveencephalopathy, hemorrhagic disorders, and disorders involving aberrantfunction of the blood-brain barrier (e.g., CNS infections such asmeningitis and encephalitis, aseptic meningitis, metastasis of non-CNStumor cells into the CNS, various pain disorders such as migraine,blindness and other vision problems, and CNS-related adverse drugreactions such as head pain, sleepiness, and confusion). TANGO 273proteins, nucleic acids encoding them, and agents that modulate activityor expression of either of these can be used to prognosticate, diagnose,and treat one or more of these disorders.

[0212] Developmental regulation of TANGO 273 expression in fetalneurological tissues, as described herein, is an indication that TANGO273 proteins, nucleic acids, and modulators thereof can be used toprognosticate, diagnose, and treat one or more disorders which involveaberrant fetal neurological development. Examples of such disordersinclude blindness, deafness, fetal death, mental retardation, dysraphia,anencephaly, malformation of cerebral hemispheres, encephalocele,porencephaly, hydranencephaly, hydrocephalus, and spina bifida.

[0213] The fact that TANGO 273 is expressed in tissues which wereexposed to LPS indicates that TANGO 273 mediates one or morephysiological responses of cells to bacterial infection. Thus, TANGO 273is involved in one or more of detection of bacteria in a tissue in whichit is expressed, movement of cells with relation to sites of bacterialinfection, production of biological molecules which inhibit bacterialinfection, and production of biological molecules which alleviatecellular or other physiological damage wrought by bacterial infection.

[0214] Presence in TANGO 273 protein of multiple SH3 domain bindingsites indicates that TANGO 273 protein interacts with one or more SH3domain- containing proteins. Thus, TANGO 273 protein mediates binding ofproteins (i.e., binding of proteins to TANGO 273 and to one another toform protein complexes) in cells in which it is expressed. TANGO 273 isalso involved in transduction of signals between the exteriorenvironment of cells (i.e., including from other cells) and the interiorof cells in which it is expressed. TANGO 273 mediates regulation of cellgrowth and proliferation, endocytosis, activation of respiratory burst,and other physiological processes triggered by transmission of a signalvia a protein with which TANGO 273 interacts.

[0215] Sequence similarity of TANGO 273 cDNA with an EST expressed inprostate tissue indicates that TANGO 273 can be expressed in prostatetissue, and can thus be involved in disorders of the prostate. Thus,TANGO 273 proteins, nucleic acids encoding them, and agents thatmodulate activity or expression of either of these can be used to treatprostate disorders. Examples of prostate disorders which can be treatedin this manner include inflammatory prostatic diseases (e.g., acute andchronic prostatitis and granulomatous prostatitis), prostatichyperplasia (e.g., benign prostatic hypertrophy or hyperplasia), andprostate tumors (e.g., carcinomas).

[0216] In another example, TANGO 273 polypeptides, nucleic acids, ormodulators thereof, can be used to treat cardiovascular disorders, suchas ischemic heart disease (e.g., angina pectoris, myocardial infarction,and chronic ischemic heart disease), hypertensive heart disease,pulmonary heart disease, valvular heart disease (e.g., rheumatic feverand rheumatic heart disease, endocarditis, mitral valve prolapse, andaortic valve stenosis), congenital heart disease (e.g., valvular andvascular obstructive lesions, atrial or ventricular septal defect, andpatent ductus arteriosus), or myocardial disease (e.g., myocarditis,congestive cardiomyopathy, and hypertrophic cardiomyopathy).

[0217] In another example, TANGO 273 polypeptides, nucleic acids, ormodulators thereof, can be used to treat disorders of the brain, such ascerebral edema, hydrocephalus, brain herniations, iatrogenic disease(due to, e.g., infection, toxins, or drugs), inflammations (e.g.,bacterial and viral meningitis, encephalitis, and cerebraltoxoplasmosis), cerebrovascular diseases (e.g., hypoxia, ischemia, andinfarction, intracranial hemorrhage and vascular malformations, andhypertensive encephalopathy), and tumors (e.g., neuroglial tumors,neuronal tumors, tumors of pineal cells, meningeal tumors, primary andsecondary lymphomas, intracranial tumors, and medulloblastoma), and totreat injury or trauma to the brain.

[0218] In another example, TANGO 273 polypeptides, nucleic acids, ormodulators thereof, can be used to treat disorders of skeletal muscle,such as muscular dystrophy (e.g., Duchenne muscular dystrophy, Beckermuscular dystrophy, Emery-Dreifuss muscular dystrophy, limb-girdlemuscular dystrophy, facioscapulohumeral muscular dystrophy, myotonicdystrophy, oculopharyngeal muscular dystrophy, distal musculardystrophy, and congenital muscular dystrophy), motor neuron diseases(e.g., amyotrophic lateral sclerosis, infantile progressive spinalmuscular atrophy, intermediate spinal muscular atrophy, spinal bulbarmuscular atrophy, and adult spinal muscular atrophy), myopathies (e.g.,inflammatory myopathies such as dermatomyositis and polymyositis,myotonia congenita, paramyotonia congenita, central core disease,nemaline myopathy, myotubular myopathy, and periodic paralysis), andmetabolic diseases of muscle (e.g., phosphorylase deficiency, acidmaltase deficiency, phosphofructokinase deficiency, debrancher enzymedeficiency, mitochondrial myopathy, camitine deficiency, camitinepalmityl transferase deficiency, phosphoglycerate kinase deficiency,phosphoglycerate mutase deficiency, lactate dehydrogenase deficiency,and myoadenylate deaminase deficiency).

[0219] In another example, TANGO 273 polypeptides, nucleic acids, ormodulators thereof, can be used to treat pancreatic disorders, such aspancreatitis (e.g., acute hemorrhagic pancreatitis and chronicpancreatitis), pancreatic cysts (e.g., congenital cysts, pseudocysts,and benign or malignant neoplastic cysts), pancreatic tumors (e.g.,pancreatic carcinoma and adenoma), diabetes mellitus (e.g., insulin- andnon-insulin-dependent types, impaired glucose tolerance, and gestationaldiabetes), or islet cell tumors (e.g., insulinomas, adenomas,Zollinger-Ellison syndrome, glucagonomas, and somatostatinoma).

[0220] In another example, TANGO 273 polypeptides, nucleic acids, ormodulators thereof, can be used to treat placental disorders, such astoxemia of pregnancy (e.g., preeclampsia and eclampsia), placentitis, orspontaneous abortion.

[0221] In another example, TANGO 273 polypeptides, nucleic acids, ormodulators thereof, can be used to treat pulmonary disorders, such asatelectasis, cystic fibrosis, rheumatoid lung disease, pulmonarycongestion or edema, chronic obstructive airway disease (e.g.,emphysema, chronic bronchitis, bronchial asthma, and bronchiectasis),diffuse interstitial diseases (e.g., sarcoidosis, pneumoconiosis,hypersensitivity pneumonitis, Goodpasture's syndrome, idiopathicpulmonary hemosiderosis, pulmonary alveolar proteinosis, desquamativeinterstitial pneumonitis, chronic interstitial pneumonia, fibrosingalveolitis, hamman-rich syndrome, pulmonary eosinophilia, diffuseinterstitial fibrosis, Wegener's granulomatosis, lymphomatoidgranulomatosis, and lipid pneumonia), or tumors (e.g., bronchogeniccarcinoma, bronchioalveolar carcinoma, bronchial carcinoid, hamartoma,and mesenchymal tumors).

[0222] In another example, TANGO 273 polypeptides, nucleic acids, ormodulators thereof, can be used to treat hepatic (liver) disorders, suchas jaundice, hepatic failure, hereditary hyperbilirubinemias (e.g.,Gilbert's syndrome, Crigler-Naijar syndromes, and Dubin-Johnson andRotor's syndromes), hepatic circulatory disorders (e.g., hepatic veinthrombosis and portal vein obstruction and thrombosis) hepatitis (e.g.,chronic active hepatitis, acute viral hepatitis, and toxic anddrug-induced hepatitis) cirrhosis (e.g., alcoholic cirrhosis, biliarycirrhosis, and hemochromatosis), or malignant tumors (e.g., primarycarcinoma, hepatoblastoma, and angiosarcoma).

[0223] In another example, TANGO 273 polypeptides, nucleic acids, ormodulators thereof, can be used to treat renal (kidney) disorders, suchas glomerular diseases (e.g., acute and chronic glomerulonephritis,rapidly progressive glomerulonephritis, nephrotic syndrome, focalproliferative glomerulonephritis, glomerular lesions associated withsystemic disease such as systemic lupus erythematosus, Goodpasture'ssyndrome, multiple myeloma, diabetes, neoplasia, sickle cell disease,and chronic inflammatory diseases), tubular diseases (e.g., acutetubular necrosis and acute renal failure, polycystic renal disease,medullary sponge kidney, medullary cystic disease, nephrogenic diabetes,and renal tubular acidosis), tubulointerstitial diseases (e.g.,pyelonephritis, drug and toxin induced tubulointerstitial nephritis,hypercalcemic nephropathy, and hypokalemic nephropathy) acute andrapidly progressive renal failure, chronic renal failure,nephrolithiasis, vascular diseases (e.g., hypertension andnephrosclerosis, microangiopathic hemolytic anemia, atheroembolic renaldisease, diffuse cortical necrosis, and renal infarcts), or tumors(e.g., renal cell carcinoma and nephroblastoma).

[0224] TANGO 286

[0225] A cDNA clone (designated jthkf042e03) encoding at least a portionof human TANGO 286 protein was isolated from a human keratinocyte cDNAlibrary. The human TANGO 286 protein is predicted by structural analysisto be a secreted protein.

[0226] The full length of the cDNA encoding TANGO 286 protein (FIG. 5;SEQ ID NO: 33) is 1980 nucleotide residues. The ORF of this cDNA,nucleotide residues 133 to 1497 of SEQ ID NO: 33 (i.e., SEQ ID NO: 34),encodes a 455-amino acid secreted protein (FIG. 5; SEQ ID NO: 35).

[0227] The invention thus includes purified TANGO 286 protein, both inthe form of the immature 455 amino acid residue protein (SEQ ID NO: 35)and in the form of the mature 432 amino acid residue protein (SEQ ID NO:37). Mature TANGO 286 protein can be synthesized without the signalsequence polypeptide at the amino terminus thereof, or it can besynthesized by generating immature TANGO 286 protein and cleaving thesignal sequence therefrom.

[0228] In addition to full length mature and immature TANGO 286proteins, the invention includes fragments, derivatives, and variants ofthese TANGO 286 proteins, as described herein. These proteins,fragments, derivatives, and variants are collectively referred to hereinas polypeptides of the invention or proteins of the invention.

[0229] The invention also includes nucleic acid molecules which encode apolypeptide of the invention. Such nucleic acids include, for example, aDNA molecule having the nucleotide sequence listed in SEQ ID NO: 33 orsome portion thereof, such as the portion which encodes mature TANGO 286protein, immature TANGO 286 protein, or a domain of TANGO 286 protein.These nucleic acids are collectively referred to as nucleic acids of theinvention.

[0230] TANGO 286 proteins and nucleic acid molecules encoding themcomprise a family of molecules having certain conserved structural andfunctional features.

[0231] A common domain of TANGO 286 proteins is a signal sequence. Asused herein, a signal sequence includes a peptide of at least about 10amino acid residues in length which occurs at the amino terminus ofmembrane-bound proteins and which contains at least about 45%hydrophobic amino acid residues such as alanine, leucine, isoleucine,phenylalanine, proline, tyrosine, tryptophan, or valine. In a preferredembodiment, a signal sequence contains at least about 10 to 35 aminoacid residues, preferably about 10 to 20 amino acid residues, and has atleast about 35-60%, more preferably 40-50%, and more preferably at leastabout 45% hydrophobic residues. A signal sequence serves to direct aprotein containing such a sequence to a lipid bilayer. Thus, in oneembodiment, a TANGO 286 protein contains a signal sequence correspondingto amino acid residues 1 to 23 of SEQ ID NO: 35 (SEQ ID NO: 36). Thesignal sequence is cleaved during processing of the mature protein.

[0232] TANGO 286 is a secreted soluble protein (i.e., a secreted proteinhaving a single extracellular domain), as indicated by computerizedsequence analysis and comparison of the amino acid sequence of TANGO 286with related proteins, such as the soluble proteins designatedbactericidal permeability increasing (BPI) protein and recombinantendotoxin neutralizing polypeptide (RENP).

[0233] TANGO 286 proteins typically comprise a variety of potentialpost-translational modification sites (often within an extracellulardomain), such as those described herein in Table IX, as predicted bycomputerized sequence analysis of TANGO 286 proteins using amino acidsequence comparison software (comparing the amino acid sequence of TANGO286 with the information in the PROSITE database {rel. 12.2; February,1995} and the Hidden Markov Models database {Rel. PFAM 3.3 }). Incertain embodiments, a protein of the invention has at least 1, 2, 4, 6,10, 15, or 20 or more of the post-translational modification siteslisted in Table IX. TABLE IX Amino Acid Type of Potential ModificationSite or Residues of Amino Acid Domain SEQ ID NO: 35 SequenceN-glycosylation site 79 to 82 NFSN 92 to 95 NTSL 113 to 116 NIST 161 to164 NLST 173 to 176 NYTL 205 to 208 NLTD 249 to 252 NLTL 303 to 306 NFTL320 to 323 NSTV 363 to 366 NRSN Protein kinase C phosphorylation site 35to 37 TQR 362 to 364 SNR 429 to 431 SSK Casein kinase II phosphorylationsite 63 to 66 SGSE 130 to 133 SFAE 163 to 166 STLE 169 to 172 TKID 175to 178 TLLD 183 to 186 SSPE 253 to 256 STEE 321 to 324 STVE 365 to 368SNIE 409 to 412 SDIE N-myristoylation site 42 to 47 GVQAGM 269 to 274GNVLSR Lipid-binding serum glycoprotein  12 to 427 see FIG. 5 domain

[0234] Certain lipid-binding serum glycoproteins, such as LPS-bindingprotein (LBP), bactericidal permeability-increasing protein (BPI),cholesteryl ester transfer protein (CETP), and phospholipid transferprotein (PLTP), share regions of sequence similarity which are hereindesignated a lipid-binding serum glycoprotein domain (Schumann et al.,(1990) Science 249:1429-1431; Gray et al., (1989) J. Biol. Chem.264:9505-9509; Day et al., (1994) J. Biol. Chem. 269:9388-9391). Theconsensus pattern of lipid-binding serum glycoprotein domains is asfollows (using standard single letter amino acid abbreviations wherein Xis any amino acid residue).

-(P or A)-(G or A)-(L or I or V or M or C)-X₂-R-(I or V)-(S orT)-X₃-L-X_((4 or 5))-(E or Q)-X₄-(L or I or V or M)-X_((0 or 1))-(E or Qor K)-X₈-P-(e.g., amino acid residues 28-60 of SEQ ID NO: 35).

[0235] Proteins in which a lipid-binding serum glycoprotein domainoccurs are often structurally related and exhibit related physiologicalactivities. LBP binds to lipid A moieties of bacterial LPS and, oncebound thereto, induces secretion of a-tumor necrosis factor, apparentlyby interacting with the CD14 receptor. BPI also binds LPS and exerts acytotoxic effect on Gram-negative bacteria (Elsbach, (1998) J. Leukoc.Biol. 64:14-18). CETP is involved in transfer of insoluble cholesterylesters during reverse cholesterol transport. PLTP appears to be involvedin phospholipid transport and modulation of serum HDL particles.

[0236] The signal peptide prediction program SIGNALP (Nielsen et al.(1997) Protein Engineering 10:1-6) predicted that TANGO 286 proteinincludes a 23 amino acid signal peptide (amino acid residues 1 to 23 ofSEQ ID NO: 35; SEQ ID NO: 36) preceding the mature TANGO 286 protein(amino acid residues 24 to 455 of SEQ ID NO: 35; SEQ ID NO: 37). HumanTANGO 286 protein is a secreted soluble protein.

[0237]FIG. 5E depicts a hydrophilicity plot of TANGO 286 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Asdescribed elsewhere herein, relatively hydrophilic regions are generallylocated at or near the surface of a protein, and are more frequentlyeffective immunogenic epitopes than are relatively hydrophobic regionsFor example, the region of human TANGO 286 protein from about amino acidresidue 420 to about amino acid residue 435 appears to be located at ornear the surface of the protein, while the region from about amino acidresidue 325 to about amino acid residue 345 appears not to be located ator near the surface.

[0238] The predicted molecular weight of TANGO 286 protein withoutmodification and prior to cleavage of the signal sequence is about 50.9kilodaltons. The predicted molecular weight of the mature TANGO 286protein without modification and after cleavage of the signal sequenceis about 48.2 kilodaltons.

[0239] The gene encoding human TANGO 286 protein was determined to belocated on chromosome 22 by comparison of matching genomic clones suchas the clones assigned GenBank Accession numbers W16806 and AL021937.

[0240] A portion of TANGO 286 protein exhibits significant amino acidhomology with a region of the human chromosome region 22q12-13 genomicnucleotide sequence having GenBank Accession number AL021937. Alignmentof a 45 kilobase nucleotide sequence encoding TANGO 286 with AL021937,however, indicated the presence in TANGO 286 of exons which differ fromthose disclosed in L021937 (pam120.mat scoring matrix; gap penalties−12/−4). This region of chromosome 22 comprises an immunoglobulin lambdachain C (IGLC) pseudogene, the Ret finger protein-like 3 (RFPL3) and Retfinger protein-like 3 antisense (RFPL3S) genes, a gene encoding a novelimmunoglobulin lambda chain V family protein, a novel gene encoding aprotein similar both to mouse RGDS protein (RALGDS, RALGEF, guaninenucleotide dissociation stimulator A) and to rabbit oncogene RSC, anovel gene encoding the human orthologue of worm F16A11.2 protein, anovel gene encoding a protein similar both to BPI and to rabbitliposaccharide-binding protein, and a 5′-portion of a novel gene. Thisregion also comprises various ESTs, STSs, GSSs, genomic marker D22S1175,a ca repeat polymorphism and putative CpG islands. TANGO 286 proteinthus shares one or more structural or functional features of thesemolecules.

[0241] TANGO 286 protein exhibits considerable sequence similarity withBPI protein, having 23.9% amino acid sequence identity therewith, asassessed using the ALIGN v. 2.0 computer software using a pam120.matscoring matrix and gap penalties of −12/−4. TANGO 286 protein alsoexhibits considerable sequence similarity with recombinant endotoxinneutralizing polypeptide (RENP), having 24.5% amino acid sequenceidentity therewith, as assessed using the ALIGN software. Physiologicalactivities of BPI protein and RENP have been described (e.g., Gabay etal., (1989) Proc. Natl. Acad. Sci. USA 86:5610-5614; Elsbach, (1998) J.Leukoc. Biol. 64:14-18; Mahadeva et al., (1997) Chest 112:1699-1701;International patent application WO96/34873). RENP, for example, bindsLPS and neutralizes bacterial endotoxins. BPI, RENP, and other proteinsin which a lipid-binding serum glycoprotein domain occurs bind LPS andneutralize bacterial endotoxins, and are therefore useful forpreventing, detecting, and treating LPS-related disorders such as shock,disseminated intravascular coagulation, anemia, thrombocytopenia, adultrespiratory distress syndrome, renal failure, liver disease, anddisorders associated with Gram negative bacterial infections. Inaddition to the physiological conditions described above, BPI protein isknown to be involved in vasculitis and bronchiectasis, in thatantibodies which bind specifically with BPI protein are present in atleast some patients afflicted with these disorders (Mahadeva et al.,supra).

[0242] Biological Function of TANGO 286 Proteins, Nucleic Acids, andModulators Thereof

[0243] Expression of TANGO 286 in keratinocyte library indicates thatthis protein is involved in a disorders which involve keratinocytes.Such disorders include, for example, disorders involving extracellularmatrix abnormalities, dermatological disorders, ocular disorders,inappropriate hair growth (e.g., baldness), infections of the nails ofthe fingers and toes, scalp disorders (e.g., dandruff), and the like.

[0244] The fact that TANGO 286 protein contains a lipid-binding serumglycoprotein domain indicates that TANGO 286 is involved in one or morephysiological processes in which these other lipid-binding serumglycoprotein domain-containing proteins are involved. Thus, TANGO 286 isinvolved in one or more of lipid transport, metabolism, serum lipidparticle regulation, host anti-microbial defensive mechanisms, and thelike.

[0245] Human TANGO 286 shares physiological functionality with otherproteins in which a lipid-binding serum glycoprotein domains occurs(e.g., LBP, BPI protein, CETP, and PLTP). Based on the amino acidsequence similarity of TANGO 286 with BPI protein and with RENP, TANGO286 protein exhibits physiological activities exhibited by theseproteins. Thus, TANGO 286 proteins are useful for preventing,diagnosing, and treating, among others, lipid transport disorders, lipidmetabolism disorders, disorders of serum lipid particle regulation,obesity, disorders involving insufficient or inappropriate hostanti-microbial defensive mechanisms, vasculitis, bronchiectasis,LPS-related disorders such as shock, disseminated intravascularcoagulation, anemia, thrombocytopenia, adult respiratory distresssyndrome, renal failure, liver disease, and disorders associated withGram negative bacterial infections, such as bacteremia, endotoxemia,sepsis, and the like.

[0246] TANGO 294

[0247] A cDNA clone (designated jthrc145g07) encoding at least a portionof human TANGO 294 protein was isolated from a human pulmonary arterysmooth muscle cell cDNA library. The human TANGO 294 protein ispredicted by structural analysis to be a transmembrane membrane protein.Expression of DNA encoding TANGO 294 was observed, using a variety ofmethods, in pancreas, lung tumor, stomach, pulmonary artery smoothmuscle cells, and colon tumor tissues and in activated peripheral bloodmononuclear cells (PBMCs). More detailed expression data is describedherein.

[0248] The full length of the cDNA encoding TANGO 294 protein (FIG. 6;SEQ ID NO: 45) is 2044 nucleotide residues. The ORF of this cDNA,nucleotide residues 126 to 1394 of SEQ ID NO: 45 (i.e., SEQ ID NO: 46),encodes a 423-amino acid transmembrane protein (FIG. 6; SEQ ID NO: 47).

[0249] The invention includes purified TANGO 294 protein, both in theform of the immature 423 amino acid residue protein (SEQ ID NO: 47) andin the form of the mature 390 amino acid residue protein (SEQ ID NO:49). Mature TANGO 294 protein can be synthesized without the signalsequence polypeptide at the amino terminus thereof, or it can besynthesized by generating immature TANGO 294 protein and cleaving thesignal sequence therefrom.

[0250] In addition to full length mature and immature TANGO 294proteins, the invention includes fragments, derivatives, and variants ofTANGO 294 protein, as described herein. These proteins, fragments,derivatives, and variants are collectively referred to herein aspolypeptides of the invention or proteins of the invention.

[0251] The invention also includes nucleic acid molecules which encode apolypeptide of the invention. Such nucleic acids include, for example, aDNA molecule having the nucleotide sequence listed in SEQ ID NO: 45 orsome portion thereof, such as the portion which encodes mature TANGO 294protein, immature TANGO 294 protein, or a domain of TANGO 294 protein.These nucleic acids are collectively referred to as nucleic acids of theinvention.

[0252] TANGO 294 proteins and nucleic acid molecules encoding themcomprise a family of molecules having certain conserved structural andfunctional features.

[0253] Also included within the scope of the invention are TANGO 294proteins having a signal sequence. As used herein, a signal sequenceincludes a peptide of at least about 10 amino acid residues in lengthwhich occurs at the amino terminus of membrane-bound proteins and whichcontains at least about 45% hydrophobic amino acid residues such asalanine, leucine, isoleucine, phenylalanine, proline, tyrosine,tryptophan, or valine. In a preferred embodiment, a signal sequencecontains at least about 10 to 35 amino acid residues, preferably about10 to 20 amino acid residues, and has at least about 35-60%, morepreferably 40-50%, and more preferably at least about 45% hydrophobicresidues. A signal sequence serves to direct a protein containing such asequence to a lipid bilayer. Thus, in one embodiment, a TANGO 294protein contains a signal sequence corresponding to amino acid residues1 to 33 of SEQ ID NO: 47 (SEQ ID NO: 48). The signal sequence is cleavedduring processing of the mature protein.

[0254] The naturally-occurring form of TANGO 294 protein is a secretedprotein (i.e., not comprising the predicted signal sequence). However,in variant forms, TANGO 294 proteins can be transmembrane proteins whichinclude an extracellular domain. In this transmembrane variant form, thepredicted TANGO 294 protein extracellular domain is located from aboutamino acid residue 34 to about amino acid residue 254 of SEQ ID NO: 47,the predicted cytoplasmic domain is located from about amino acidresidue 280 to amino acid residue 423 of SEQ ID NO: 47 (SEQ ID NO: 52),and the predicted transmembrane domain is located from about amino acidresidues 255 to 279 of SEQ ID NO: 47 (SEQ ID NO: 51).

[0255] TANGO 294 proteins typically comprise a variety of potentialpost-translational modification sites (often within an extracellulardomain), such as those described herein in Table X, as predicted bycomputerized sequence analysis of TANGO 294 proteins using amino acidsequence comparison software (comparing the amino acid sequence of TANGO294 with the information in the PROSITE database {rel. 12.2; February,1995} and the Hidden Markov Models database {Rel. PFAM 3.3}). In certainembodiments, a protein of the invention has at least 1, 2, 4, 6, 10, 15,or 20 or more of the post-translational modification sites listed inTable X. TABLE X Amino Acid Type of Potential Modification Site Residuesof Amino Acid or Domain SEQ ID NO: 47 Sequence N-glycosylation site 48to 51 NISE 113 to 116 NNSL 285 to 288 NMSR 413 to 416 NLSQ Proteinkinase C phosphorylation 12 to 14 SHR site 138 to 140 SRK 217 to 219 TVKCasein kinase II phosphorylation 155 to 158 SYDE site 175 to 178 TGQE198 to 201 TMPE 360 to 363 SNPE Tyrosine kinase phosphorylation 174 to182 KTGQEKIYY site N-myristoylation site  99 to 104 GLVGGA 130 to 135GNSRGN 188 to 193 GTTMGF 277 to 282 GGFNTN Amidation site 240 to 243FGKK Lipase serine active site 180 to 189 IYYVGYSQGT Lipase ConservedActive Site 186 5 Residues 357 D 386 H Lipase Conserved Cysteine 260 CResidues 269 C Lipase Conserved Oxyanion Hole 100 L Residues 187 QAlpha/beta hydrolase fold domain 125 to 404 See FIG. 6

[0256] Alpha/beta hydrolase fold domains occur in a wide variety ofenzymes (Ollis et al., (1992) Protein Eng. 5:197-211). The alpha/betafold domain is a conserved topological domain in which sequence homologyis not necessarily conserved. Conservation of topology in the alpha/betafold domain preserves arrangement of catalytic residues, even thoughthose residues, and the reactions they catalyze, can vary. In manyenzymes, particularly including alpha/beta hydrolases, this domainencompasses the active site of the enzyme. In one embodiment, theprotein of the invention has at least one domain that is at least 55%,preferably at least about 65%, more preferably at least about 75%, yetmore preferably at least about 85%, and most preferably at least about95% identical to the alpha/beta hydrolase fold domain described hereinin Table X.

[0257] The signal peptide prediction program SIGNALP (Nielsen et al.(1997) Protein Engineering 10:1-6) predicted that human TANGO 294protein includes a 33 amino acid signal peptide (amino acid residues 1to 33 of SEQ ID NO: 47; SEQ ID NO: 48) preceding the mature TANGO 294protein (amino acid residues 34 to 423 of SEQ ID NO: 47; SEQ ID NO: 49).Human TANGO 294 protein is a soluble secreted protein. However, in thetransmembrane variant form, human TANGO 294 protein includes anextracellular domain (amino acid residues 34 to 254 of SEQ ID NO: 47;SEQ ID NO: 50); a transmembrane domain (amino acid residues 255 to 279of SEQ ID NO: 47; SEQ ID NO: 51); and a cytoplasmic domain (amino acidresidues 280 to 423 of SEQ ID NO: 47; SEQ ID NO: 52).

[0258]FIG. 6F depicts a hydrophilicity plot of human TANGO 294 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Thehydrophobic region which corresponds to amino acid residues 1 to 33 ofSEQ ID NO: 47 is the signal sequence of human TANGO 294 (SEQ ID NO: 49).The hydrophobic region which corresponds to amino acid residues 255 to279 of SEQ ID NO: 47 is the predicted transmembrane domain of humanTANGO 294 (SEQ ID NO: 51). As described elsewhere herein, relativelyhydrophilic regions are generally located at or near the surface of aprotein, and are more frequently effective immunogenic epitopes than arerelatively hydrophobic regions. For example, the region of human TANGO294 protein from about amino acid residue 130 to about amino acidresidue 150 appears to be located at or near the surface of the protein,while the region from about amino acid residue 90 to about amino acidresidue 100 appears not to be located at or near the surface.

[0259] The predicted molecular weight of human TANGO 294 protein withoutmodification and prior to cleavage of the signal sequence is about 48.2kilodaltons. The predicted molecular weight of the mature human TANGO294 protein without modification and after cleavage of the signalsequence is about 44.2 kilodaltons.

[0260] It may be that amino acid residues 1 to 15 of SEQ ID NO: 47 donot occur in TANGO 294 protein. However, it is recognized that aminoacid residues 16 to 33 of SEQ ID NO: 47 form a functional signalsequence even in the absence of residues 1 to 15. The amino acidsequence (and hence the properties) of mature TANGO 294 protein areunaffected by presence or absence of amino acid residues 1 to 15 ofimmature TANGO 294 protein.

[0261] Human TANGO 294 protein exhibits considerable sequence similarity(i.e., about 75% amino acid sequence identity) to lingual and gastriclipase proteins of rat (Swissprot Accession no. P04634; Docherty et al.(1985) Nucleic Acids Res. 13:1891-1903), dog (Swissprot Accession no.P80035; Carriere et al. (1991) Eur. J. Biochem. 202:75-83), and human(Swissprot Accession no. P07098; Bembaeck and Blaeckberg (1987) Biochim.Biophys. Acta 909:237-244), as assessed using the ALIGN v. 2.0 computersoftware using a pam12.mat scoring matrix and gap penalties of −12/−4.TANGO 294 is distinct from the known human lipase, as indicated in FIGS.6D and 6E. FIGS. 6D and 6E depict an alignment of the amino acidsequences of human TANGO 294 protein (SEQ ID NO: 47) and the known humanlipase protein (SEQ ID NO: 75), as assessed using the same software andparameters. In this alignment (pam120.mat scoring matrix, gap penalties−12/−4), the amino acid sequences of the proteins are 49.8% identical.TANGO 294 also is distinct from the known human lysosomal acid lipase,as indicated in FIGS. 6G and 6H. FIGS. 6G and 6H depicts an alignment ofthe amino acid sequences of human TANGO 294 protein (SEQ ID NO: 47) andthe known human lysosomal acid lipase protein (SEQ ID NO: 41). In thisalignment (pam120.mat scoring matrix, gap penalties −12/−4), the aminoacid sequences of the proteins are 56.9% identical.

[0262] TANGO 294 is a human lipase distinct from the known human lipaseand the known human lysosomal acid lipase. Furthermore, in view of thecomparisons of the amino acid sequences of TANGO 294 and the two humanlipases and the nature of transcriptional initiation sites, it isrecognized that the transcriptional start site can correspond to eitherof the methionine residues located at residues 1 and 15 of SEQ ID NO: 47The present invention thus includes proteins in which the initiallytranscribed amino acid residue is the methionine residue at position 1of SEQ ID NO: 47 and proteins in which the initially transcribed aminoacid residue is the methionine residue at position 15 of SEQ ID NO: 47(i.e., proteins in which the amino acid sequence of TANGO 294 does notinclude residues 1 to 14 of SEQ ID NO: 47). Furthermore, because aminoacid residues 1 to 14 of SEQ ID NO: 47 are predicted to be part of asignal sequence, it is recognized that the protein not comprising thisportion of the amino acid sequence will nonetheless exhibit a functionalsignal sequence at its amino terminus.

[0263] TANGO 294 Expression Analysis

[0264] TaqMan™ Experiments

[0265] Total RNA was prepared from various human tissues by a singlestep extraction method using RNA STAT-60 according to the manufacturer'sinstructions (TelTest, Inc). Each RNA preparation was treated with DNaseI (Ambion) at 37° C. for 1 hour. DNAse I treatment was determined to becomplete if the sample required at least 38 PCR amplification cycles toreach a threshold level of fluorescence using beta-2 microglobulin as aninternal amplicon reference. The integrity of the RNA samples followingDNase I treatment was confirmed by agarose gel electrophoresis andethidium bromide staining. After phenol extraction cDNA was preparedfrom the sample using the SUPERSCRIPT™ Choice System following themanufacturer's instructions (Gibco BRL). A negative control of RNAwithout reverse transcriptase was mock reverse transcribed for each RNAsample.

[0266] Novel TANGO 294 expression was measured by TaqMan™ quantitativePCR (Perkin Elmer Applied Biosystems) in cDNA prepared from thefollowing human tissues:

[0267] Tissue Panel One (Phase I, General Expression in normal andtumorigenic tissues): Artery normal, Aorta diseased, Vein normal,Coronary SMC, HUVEC, Hemangioma, Heart (normal), Congestive HeartFailure, Kidney, Skeletal Muscle, Adipose (normal), Pancreas, PrimaryOsteoblasts, Osteoclasts (differentiated), Skin (normal), Spinal Cord(normal), Brain Cortex (normal), Brain Hypothalamus (normal), Nerve,Dorsal Root Ganglia, Breast (normal), Breast Tumor, Ovary (normal),Ovary Tumor, Prostate (normal), Prostate Tumor, Salivary Glands, Colon(pools of 3 normal colon tissues), Colon Tumor (3 colonadenocarcinomas), Lung (normal), Lung Tumor, Lung (COPD), Colon (3 colonIBD samples), Liver (normal), Liver Fibrosis, Spleen (normal), Tonsil(normal), Lymph Node (normal), Small Intestine (normal), Macrophages,Synovium, Bone Marrow-MNC, Activated Peripheral Blood Mononuclear Cells(PBMC), Neutrophils, Megakaryocytes, and Erthyroid.

[0268] Tissue Panel Two (Phase II, General Expression Pattern in SolidTumors): Breast (normal, 3 separate samples), Breast (tumor, 5 separatesamples), Lymph node (metastasized from breast), Lung (metastasized frombreast), Ovary (normal, 2 separate samples), Ovary (tumor, 5 separatesamples), Lung (normal, 3 separate samples), Lung (tumor, 6 separatesamples), Colon (normal, 3 separate samples), Colon (Adenocarcinoma, 4separate samples), Liver (metastasized from colon, 3 separate samples),Liver (normal, female), Cervix Squamous Carcinoma (2 separate samples),Human Microvascular Endothelial Cells- Arr, Human MicrovascularEndothelial Cells- Prol, Pooled Hemangiomas, HCT116N22 Normoxic, andHCT116H22 Hypoxic.

[0269] Tissue Panel Three (Colon Cancer Expanded Panel: Specificexpression pattern in various stages of colorectal cancer): Colon(normal, 6 separate samples), Colon (adenomas, 2 separate samples),Colon (stage B adenocarcinomas, 6 separate samples), Colon (stage Cadenocarcinomas, 6 separate samples), Liver (normal, 6 separatesamples), Liver (metastasized from colon, 6 separate samples), and 1abdominal colon metastasis sample.

[0270] Tissue Panel Four (Xenograft Panel): MCF-7 Breast T, ZR75 BreastT, T47D Breast T, MDA 231 Breast T, MDA 435 Breast T, SKBr3 Breast, DLD1 ColonT (stage C), SW480 Colon T (stage B), SW620 ColonT (stage C),HCT116, HT29, Colon 205, NCIH125, NCIH67, NCIH322, NCIH460, A549, NHBE,SKOV-3 ovary, OVCAR-3 ovary, 293 Baby Kidney, and 293T Baby Kidney.

[0271] Tissue Panel Five (Colon to Liver Metastases Panel: Specificexpression patterns in late stage colon cancer (metastasis)): Colon(normal, 3 separate samples), Colonic ACA-C, Colonic ACA-C, ColonicACA-B, Colon (adenocarcinoma), Liver (metastasized from colon, 17separate samples), and Liver (normal, 3 separate samples).

[0272] Probes were designed by PrimerExpress software (PE Biosystems)based on the sequence of each gene. Each gene probe was labeled usingFAM (6-carboxyfluorescein), and the beta-2 microglobulin reference probewas labeled with a different fluorescent dye, VIC. The differentiallabeling of the target gene and internal reference gene thus enabledmeasurement in same well. Forward and reverse primers and the probes forboth beta-2 microglobulin and target gene were added to the TaqMan™Universal PCR Master Mix (PE Applied Biosystems). Although the finalconcentration of primer and probe could vary, each was internallyconsistent within a given experiment. A typical experiment contained 200nanomolar of forward and reverse primers plus 100 nanomolar probe forbeta-2 microglobulin and 600 nanomolar forward and reverse primers plus200 nanomolar probe for the target gene. TaqMan matrix experiments werecarried out on an ABI PRISM 7700 Sequence Detection System (PE AppliedBiosystems). The thermal cycler conditions were as follows: hold for 2minutes at 50° C. and 10 minutes at 95° C., followed by two-step PCR for40 cycles of 95° C. for 15 seconds followed by 60° C. for 1 minute.

[0273] The following method was used to quantitatively calculate TANGO294 gene expression in the various tissues relative to beta-2microglobulin expression in the same tissue. The threshold cycle (Ct)value is defined as the cycle at which a statistically significantincrease in fluorescence is detected. A lower Ct value is indicative ofa higher mRNA concentration. The Ct value of the TANGO 294 gene isnormalized by subtracting the Ct value of the beta-2 microglobulin geneto obtain a _(Δ)Ct value using the following formula:_(Δ)Ct=Ct_(TANGO 294)-Ct_(beta-2 microglobulin). Expression is thencalibrated against a cDNA sample showing a comparatively low level ofexpression of the TANGO 294 gene. The _(Δ)Ct value for the calibratorsample is then subtracted from _(Δ)Ct for each tissue sample accordingto the following formula: _(ΔΔ)Ct=_(Δ)Ct-_(sample)-_(Δ)Ct-_(calibrator).Relative expression is then calculated using the arithmetic formulagiven by 2-^(ΔΔCt). Expression of the target TANGO 294 gene in each ofthe tissues tested is discussed in more detail below.

[0274] The TaqMan expression from Panel One shows restricted TANGO 294expression in pancreas, colon tumors, and activated peripheral bloodmononuclear cells, with highest expression in colon tumors.

[0275] The TaqMan expression from Panel Two shows TANGO 294 expressionrestricted to colon tumors and lung tumors, with a 4-50 times increasein expression in colon tumor samples over normal colon samples. Forexample, the colon tumor sample NDR 210 shows about 6× expression overnormal colon samples, the colon tumor sample CHT 382 shows about 9×expression over normal colon samples, and the colon tumor sample CHT 528shows about 30× expression over normal colon samples.

[0276] The TaqMan expression from Panel Three shows TANGO 294 elevatedexpression in adenomas and stage A, B, and C, and metastatic tumors.

[0277] The TaqMan expression from Panel Four shows TANGO 294 expressionin breast, colon, and lung cell lines (NCIH322 and NHBE).

[0278] The TaqMan expression from Panel Five shows upregulated TANGO 294expression in 80% of colon to liver metastases. Both normal colon andnormal liver show no expression.

[0279] Other TANGO 294 Expression

[0280] Transcriptional profiling data shows upregulation of TANGO 294 inearly and late stage colon tumors, as compared to adenomas and normalcolon tissue samples.

[0281] In situ hybridization experiments confirmed expression of TANGO294 in various tumor samples (restricted to a certain subset of tumorsand metastatic lesions), as well as in pancreas and inflammatory cells(e.g., PBMCs). Expression of TANGO 294 in colon tumor samples was alsoconfirmed by quantitative PCR amplification. Expression of TANGO 294 wasdetected in portions of hyperplastic colonic epithelium associated withcolonic polyps.

[0282] Biological Function of TANGO 294 proteins, nucleic acids, andmodulators thereof

[0283] The sequence similarity of TANGO 294 and mammalian lingual,gastric, and lysosomal acid lipase proteins indicates that TANGO 294 isinvolved in physiological processes identical or analogous to thoseinvolving these lipases. Lipases such as TANGO 294 catalyze formationand breakage of ester bonds between a fatty acid and a lipid moiety suchas an acylglycerol, a sterol (e.g., cholesterol), and a lipoprotein. Thelipases with which TANGO 294 exhibits the greatest similarity haveacylglycerols and sterols among their substrates, indicating that TANGO294 can exhibit preference for these substrates. Thus, TANGO 294 isinvolved in facilitating absorption and metabolism of fat. TANGO 294 canthus be used, for example, to prevent, detect, and treat disordersrelating to fat absorption and metabolism, such as inadequate expressionof gastric/pancreatic lipase, cystic fibrosis, exocrine pancreaticinsufficiency, obesity, medical treatments which alter fat absorption,and the like.

[0284] Lipases such as TANGO 294 are also involved in regulatingtransfer of fatty acid substrates such as linoleic acid and arachidonicacid among dietary lipids, intracellular lipid stores, and enzymesinvolved in biosynthesis of eicosanoids such as prostaglandins,thromboxanes, and leukotrienes. Eicosanoids are known to exert a varietyof biological effects on cells, including modulating immune responses,growth of cells, and movement of cells. Prostaglandins produced byoxidation of arachidonic acid by cyclooxygenase-2 (Cox-2) enzymes have arole in formation, growth, and spread of colon tumors (Majerus, 1998,Curr. Biol. 8(3):R87-R89). It has been hypothesized that at least partof the preventive effect of aspirin and other non-steroidalanti-inflammatory drugs on colon tumor formation is attributable to theinhibitory action of these agents on Cox-2 enzymes (Gustafson-Svard,1997, Ann. Med. 29(3):247-252). Furthermore, abnormal expression ofhormone-sensitive lipases has been observed in certain colon cancer celllines (Remaury et al., 1995, Biochem. Biophys. Res. Commun. 208(1):456),suggesting that these enzymes can have a role in colon cancerdevelopment. Taken together, these observations and the data disclosedherein indicate that TANGO 294 can modulate formation, growth, andmetastasis of colon tumors.

[0285] While not being bound by any particular theory of operation, itis believed that TANGO 294 is able to modulate the supply of eicosanoidprecursor fatty acids such as linoleic acid, gamma-linoleic acid,gamma-dihomolinoleic acid, and arachidonic acid by modulating theamounts of these acids available for eicosanoid synthesis in colontissue. These acids can be obtained by action of TANGO 294 on dietarylipids (including lipids within the colon or serum) or by esterificationor de-esterification of fatty acids and lipid stores within cells. Byway of example, TANGO 294 can catalyze formation of triacylglycerolscontaining one or more of these fatty acids, whereupon thetriacylglycerol is stored in a cellular membrane or within a lysosome.By modulating the cellular supply of these fatty acids, eicosanoidsynthesis can be modulated, and cellular processes (e.g., cellulargrowth or proliferation or movement of cells) that are affected by thepresence or absence of eicosanoids can be modulated. Thus, restrictionof the availability of eicosanoid fatty acid precursors can inhibiteicosanoid synthesis and exert a colon cancer-preventive effectanalogous to that observed by administration of a Cox-2 inhibitor suchas aspirin or sulindac.

[0286] TANGO 294 protein is known to be expressed in human pulmonaryartery smooth muscle tissue. This indicates that TANGO 294 protein isinvolved in transportation and metabolism of fats and lipids in thehuman vascular and cardiovascular systems. Thus, TANGO 294 proteins ofthe invention can be used to prevent, detect, and treat disordersinvolving these body systems. Various eicosanoids, includingthromboxanes and prostaglandins, are known to modulate vascular smoothmuscle contraction, affecting processes such as systemic blood pressureand local blood flow in tissues. The ability of TANGO 294 to modulateeicosanoid levels permits these processes to be affected. Thus,modulating expression or activity of TANGO 294 can be used to inhibit,prevent, or counteract hypo- or hypertension in a human or to modulateblood flow through a tissue in which the expression or activity ismodulated. Similarly, assessment of expression or activity of TANGO 294in a patient, or in a tissue of a patient, can predict or diagnose ablood flow or blood pressure disorder in a patient. Examples of suchdisorders include arterial hypertension, renovascular hypertension,syncope, orthostatic hypotension, and shock (including anaphylacticshock).

[0287] Certain eicosanoids are known to strongly influence epithelialand endothelial cell proliferation and the tightness of adhesion betweenepithelial or endothelial cells. By modulating eicosanoid production ina cell (e.g., a tumor cell or a PBMC), TANGO 294 can modulate theability of the cell to pass through an epithelial or endothelialmembrane. For example, many PBMCs exhibit the ability to travel withinthe bloodstream to a certain body location, at which point the cellsadhere to the vessel wall and pass through the endothelial membrane ofthe vessel in order to move to a tissue adjoining the vessel (i.e., thePBMCs exhibit the ability to extravasate). Similarly, metastatic tumorcells often exhibit an enhanced ability to move through tissue membranesand colonize body sites at which they do not normally occur (e.g., colontumor cells can metastasize to lung tissue and develop there into atumor mass). The ability of TANGO 294 to modulate these processesindicates that TANGO 294 molecules can be used to prognosticate,diagnose, inhibit, prevent, or even reverse the ability of cells toexhibit these characteristics. Thus, TANGO 294 molecules are useful inprognosticating, diagnosing, inhibiting, preventing, or reversingconditions such as inappropriate inflammation, tumor growth, and tumorcell metastasis.

[0288] Expression of TANGO 294 in tissues having an epithelial orendothelial component (e.g., colon and colon tumor, lung tumor, stomach,and pancreas tissues) indicates that TANGO 294 can influence thecohesiveness of an endothelial or epithelial membrane in one of thesetissues or the ability of cells to pass through that membrane. Theseobservations indicate that TANGO 294 can have a role in inflammatorydisorders of these and other endothelium- or epithelium-containingtissues and that TANGO 294 can affect the likelihood that a tumor cellmetastasis will enter into or take up residence within one of thesetissues. Examples of inflammatory disorders in which TANGO 294 can havea role include gastritis, ulcer, various types of colitis and other lessdefinitively characterized lower gastrointestinal disorders (e.g.,irritable and inflammatory bowel syndromes), and pancreatitis.

[0289] Expression of TANGO 294 was observed, at least at low levels, ina variety of tissues having an epithelial or endothelial portion,including colon, pancreas, small intestine, stomach, spleen, tonsil, andbreast tissues. In many such tissues where corresponding tumor tissuesamples were available (e.g., prostate, breast and lung tumor tissues),significantly greater expression of TANGO 294 was observed in the tumortissue than in the non-tumor tissue. This observation indicates thatTANGO 294 can have a role in a variety of tumors of epithelial andendothelial origin, including those in which TANGO 294 expressionoccurs. The TANGO 294 molecules described herein can therefore be usedto modulate (e.g., inhibit, prevent, alleviate, reverse, or cure)tumorigenesis and growth, proliferation, invasion, and metastasis ofthese tumors.

[0290] Expression of DNA encoding TANGO 294 is between 4 and 50 timesgreater in colon tumor tissue samples than in corresponding normal colontissue samples. Expression of TANGO 294 is elevated, relative to normalcolon tissue in colon adenomas and in colon tumors at stages A, B, andC. Elevated TANGO 294 expression is also observed in metastases ofapparent colon origin, including those which are found in liver tissue.In situ hybridization experiments confirmed expression of TANGO 294 invarious colon tumor samples, as well as in pancreas and inflammatorycells (e.g., PBMCs). Expression of TANGO 294 in colon tumor samples andcolon metastases was also confirmed by quantitative PCR amplification.Expression of TANGO 294 was detected in portions of hyperplastic colonicepithelium associated with colonic polyps.

[0291] The strong correlation described herein between expression ofTANGO 294 and occurrence of colon tumor cells in a sample indicates thatTANGO 294 is useful as a marker for detecting occurrence of colon cancerin an individual. Expression of TANGO 294 in a sample (e.g., a stoolsample, a blood sample, or a polyp or other colon biopsy) obtained froma patient can indicate a predisposition for the patient to develop acolon cancer, that a growth observed in the patient's colon is a tumor,that a colon tumor in the patient is metastasizing or has significantmetastatic potential, or that the patient has otherwise non-symptomaticcolon cancer.

[0292] The strong correlation between TANGO 294 expression and colontumor occurrence also indicates that expression of TANGO 294 can have arole in (e.g., can be used to inhibit, prevent, alleviate, reverse, orcure) one or more of tumorigenesis, colon tumor growth, or colon tumormetastasis. Compounds which modulate the activity or expression of TANGO294 (e.g., antisense oligonucleotides, antibodies which specificallybind with TANGO 294, or relatively small molecules identified byscreening) can affect the rate at which or the degree to which theseprocesses occur. Because TANGO 294 expression appears to be up-regulatedin colon tumor cells, inhibiting activity, expression, or both, of TANGO294 can inhibit or stop colon cell tumorigenesis, colon tumor growth, orcolon tumor metastasis. Expression or activity of TANGO 294 cantherefore be used in a screening assay to identify compounds which canhave one of these effects in colon cancer.

[0293] TANGO 294 is expressed in activated PBMCs. This expression isconsistent with a role for TANGO 294 in eicosanoid (e.g., leukotriene)biosynthesis. Leukotrienes are known to strongly modulate inflammationand other immune processes mediated by PBMCs. Aberrant expression ofTANGO 294 can induce PBMCs to proliferate or activate in aninappropriate manner, resulting in any of a variety of disorders.Inappropriate proliferation of PBMCs can lead to cancers such asleukemias, lymphomas, and plasma cell dyscrasias, and these cancers canbe inhibited, prevented, alleviated, reversed, or cured using modulatorsof TANGO 294 expression or activity. Inappropriate activation of PBMCscan result in a variety of inflammatory and immune disorders. By way ofexample, many autoimmune disorders involve activation of PBMCs in theabsence of a pathogen, and others involve activation of PBMCs in thepresence of a pathogen, but to an extent that immune-mediated cytotoxicactivities seriously damage one or more tissues of the patient.

[0294] Pancreatic cancer cell proliferation and survival requires asufficient level of eicosanoid synthesis (see, e.g., Ding et al., 2000,Anticancer Res. 20(4):2625-2631). Involvement of TANGO 294 in eicosanoidsynthesis and metabolism indicates that TANGO 294 can contribute toproliferation and survival of pancreatic cancer cells. TANGO 294molecules described herein can be used to inhibit, prevent, alleviate,reverse, or cure one or more of tumorigenesis, tumor cell growth, tumorcell proliferation, tumor cell invasion, and metastasis in pancreatictissue.

[0295] It has been observed that circulating fatty acids can affect theinflammatory activity of leukocytes and ameliorate inflammatorydisorders such as rheumatoid arthritis (Crocker et al., 2001, Q.J.M.94(9):475-484). Synthesis and release of eicosanoids by leukocytes canaffect the reactivity of those leukocytes and can also affect theproperties of other leukocytes and eicosanoid-sensitive tissues in thephysiological neighborhood of those leukocytes. The ability of TANGO 294to modulate synthesis and release of fatty acids in PBMCs which expressit indicates that modulation of TANGO 294 expression, activity, or both,can predict, diagnose, inhibit, or prevent inappropriate immune andinflammatory responses in patients. Thus, modulation of TANGO 294expression or activity can be used to treat patients afflicted withdisorders such as arthritis (e.g., rheumatoid arthritis), psoriasis,myasthenia gravis, dermatitis (e.g., contact dermatitis), allergies,insulin resistance, systemic lupus erythematosus, scleroderma, andautoimmune diabetes mellitus.

[0296] Involvement of TANGO 294 in activation of PBMCs indicates thatinappropriately low activation of PBMCs can be predicted, diagnosed,inhibited, prevented, or reversed by increasing expression or activityof TANGO 294 in PBMCs. Certain infectious agents (e.g., the humanimmunodeficiency virus) can deplete certain types of cells within theimmune system. Other infectious agents provoke an immune response, butthe provoked response is not sufficient to remove the infectious agentfrom the patient's system. Enhancing expression, activation, or both ofTANGO 294 in PBMCs in a patient can enhance the ability of the patient'simmune system to react to the presence of an immunogen, therebyassisting the patient in clearing the pathogen from the patient orminimizing the pathogen's impact on the patient's health.

[0297] Various eicosanoid compounds, including several thromboxanes,influence the ability of blood platelets to aggregate with one anotheror to adhere to cells of another type. The ability of TANGO 294 tomodulate production of eicosanoids, including thromboxanes, indicatesthat modulation of TANGO 294 expression, activity, or both, caninfluence the course of thrombotic disorders and disorders involvinginappropriate platelet adherence. By way of example, some thromboticdisorders (e.g., hemophilia) are characterized by insufficientthrombosis. Expression or activity of TANGO 294 can be modulated toenhance cellular production of thrombosis-enhancing eicosanoids, andassessing TANGO 294 expression or activity in a tissue can predict ordiagnose whether the patient is afflicted with a disorder characterizedby insufficient thrombosis. Conversely, assessment of TANGO 294expression or activity can be used to predict or diagnose if a patientis afflicted with (or predisposed to develop) a disorder characterizedby inappropriate thrombus formation (e.g., stroke, myocardialinfarction, or other abnormal blood coagulation) by inappropriateadherence of platelets to other tissues (e.g., coronary artery diseaseor atherosclerosis). Modulating TANGO 294 expression or activity caninhibit or prevent one of these disorders, or alleviate the disorder ifit is already occurring in a patient.

[0298] Other platelet associated disorders that TANGO 294 (or modulatorsthereof) can be used to treat, for at least the above-mentioned reasons,include, but are not limited to, thrombocytopenia due to a reducednumber of megakaryocytes in the bone marrow, for example, as a result ofchemotherapy; invasive disorders, such as leukemia, idiopathic or drug-or toxin-induced aplasia of the marrow, or rare hereditaryamegakaryocytic thrombocytopenias; ineffective thrombopoiesis, forexample, as a result of megaloblastic anemia, alcohol toxicity, vitaminB12 or folate deficiency, myelodysplastic disorders, or rare hereditarydisorders (e.g., Wiskott-Aldrich syndrome and May-hegglin anomaly); areduction in platelet distribution, for example, as a result ofcirrhosis, a splenic invasive disease (e.g., Gaucher's disease), ormyelofibrosis with extramedullary myeloid metaplasia; increased plateletdestruction, for example, as a result of removal of IgG-coated plateletsby the mononuclear phagocytic system (e.g., idiopathic thrombocytopenicpurpura (ITP), secondary immune thrombocytopenia (e.g., systemic lupuserythematosus, lymphoma, or chronic lymphocytic leukemia), drug-relatedimmune thrombocytopenias (e.g., as with quinidine, aspirin, andheparin), post-transfusion purpura, and neonatal thrombocytopenia as aresult of maternal platelet autoantibodies or maternal plateletalloantibodies). Also included are thrombocytopenia secondary tointravascular clotting and thrombin induced damage to platelets as aresult of, for example, obstetric complications, metastatic tumors,severe gram-negative bacteremia, thrombotic thrombocytopenic purpura, orsevere illness. Also included is dilutional thrombocytopenia, forexample, due to massive hemorrhage.

[0299] Platelet associated disorders also include, but are not limitedto, essential thrombocytosis and thrombocytosis associated with, forexample, splenectomy, acute or chronic inflammatory diseases, hemolyticanemia, carcinoma, Hodgkin's disease, lymphoproliferative disorders, andmalignant lymphomas.

[0300] The disorders mentioned in this section in connection with TANGO294 are collectively referred to as “TANGO 294-related disorders.”

[0301] INTERCEPT 296

[0302] A cDNA clone (designated jthEa030h09) encoding at least a portionof human INTERCEPT 296 protein was isolated from a human esophagus cDNAlibrary. The human INTERCEPT 296 protein is predicted by structuralanalysis to be a transmembrane protein having three or moretransmembrane domains. Expression of DNA encoding INTERCEPT 296 tissuehas been detected by northern analysis of human lung tissue. In humanlung tissue, two moieties corresponding to INTERCEPT 296 have beenidentified in Northern blots. It is recognized that these two moietiesmay represent alternatively polyadenylated INTERCEPT 296 mRNAs oralternatively spliced INTERCEPT 296 mRNAs. It has furthermore beenobserved that INTERCEPT 296 does not appear to be expressed in any ofheart, brain, placenta, skeletal muscle, kidney, and pancreas tissues.

[0303] The full length of the cDNA encoding INTERCEPT 296 protein (FIG.7; SEQ ID NO: 53) is 2133 nucleotide residues. The ORF of this cDNA,nucleotide residues 70 to 1098 of SEQ ID NO: 53 (i.e., SEQ ID NO: 54),encodes a 343-amino acid transmembrane protein (FIG. 7; SEQ ID NO: 55).

[0304] The invention includes purified INTERCEPT 296 protein, which hasthe amino acid sequence listed in SEQ ID NO: 55. In addition to fulllength INTERCEPT 296 proteins, the invention includes fragments,derivatives, and variants of these INTERCEPT 296 proteins, as describedherein. These proteins, fragments, derivatives, and variants arecollectively referred to herein as polypeptides of the invention orproteins of the invention.

[0305] The invention also includes nucleic acid molecules which encode apolypeptide of the invention. Such nucleic acids include, for example, aDNA molecule having the nucleotide sequence SEQ ID NO: 53 or someportion thereof, such as the portion which encodes INTERCEPT 296 proteinor a domain thereof. These nucleic acids are collectively referred to asnucleic acids of the invention.

[0306] INTERCEPT 296 proteins and nucleic acid molecules encoding themcomprise a family of molecules having certain conserved structural andfunctional features, such as the five transmembrane domains which occurin the protein.

[0307] INTERCEPT 296 comprises at least five transmembrane domains, atleast three cytoplasmic domains, and at least two extracellular domains.INTERCEPT 296 does not appear to comprise a cleavable signal sequence.Amino acid residues 1 to 70 of SEQ ID NO: 55 likely directs insertion ofthe protein into the cytoplasmic membrane. There are at least twomechanisms by which this can occur. Sequence analysis of residues 1 to70 of SEQ ID NO: 55 indicates that this entire region may represent asignal sequence or that residues 1 to 47 represent a signal sequence,with residues 48-70 representing a transmembrane region. Human INTERCEPT296 protein extracellular domains are located from about amino acidresidue 70 to about amino acid residue 182 (SEQ ID NO: 57) and fromabout amino acid residue 228 to about amino acid residue 249 (SEQ ID NO:58) of SEQ ID NO: 55. Human INTERCEPT 296 cytoplasmic domains arelocated from about amino acid residue 43 to amino acid residue 50 (SEQID NO: 64), from about amino acid residue 205 to amino acid residue 210(SEQ ID NO: 65), and from amino acid residue 272 to amino acid residue343 (SEQ ID NO: 66) of SEQ ID NO: 55. The five transmembrane domains ofINTERCEPT 296 are located from about amino acid residues 24 to 42 (SEQID NO: 59), 51 to 70 (SEQ ID NO: 60), 183 to 204 (SEQ ID NO: 61),211 to227 (SEQ ID NO: 62), and 250 to 271 (SEQ ID NO: 63) of SEQ ID NO: 55.

[0308] INTERCEPT 296 proteins typically comprise a variety of potentialpost-translational modification sites (often within an extracellulardomain), such as those described herein in Table XI, as predicted bycomputerized sequence analysis of INTERCEPT 296 proteins using aminoacid sequence comparison software (comparing the amino acid sequence ofINTERCEPT 296 with the information in the PROSITE database {rel. 12.2;February, 1995} and the Hidden Markov Models database {Rel. PFAM 3.3}).In certain embodiments, a protein of the invention has at least 1, 2, 4,6, 10, 15, or 20 or more of the post-translational modification siteslisted in Table XI. TABLE XI Amino Acid Type of Potential ModificationSite Residues of Amino Acid or Domain SEQ ID NO: 55 SequenceN-glycosylation site 71 to 74 NFSS 84 to 87 NTSY 109 to 112 NITL 121 to124 NETI 284 to 287 NQSV Protein kinase C phosphorylation 86 to 88 SYKsite 131 to 133 TWR 162 to 164 TPR 304 to 306 SPR 313 to 315 SPK 326 to328 STK Casein kinase II phosphorylation 286 to 289 SVDE site 296 to 299SPEE 309 to 312 SMAD Tyrosine kinase phosphorylation 148 to 156KGLPDPVLY site N-myristoylation site 79 to 84 GQVSTN 100 to 105 GLQVGL107 to 112 GVNITL 265 to 270 GLAMAV

[0309]FIG. 7D depicts a hydrophilicity plot of INTERCEPT 296 protein.Relatively hydrophobic regions are above the dashed horizontal line, andrelatively hydrophilic regions are below the dashed horizontal line. Thehydrophobic regions which corresponds to amino acid residues 24 to 42,51 to 70, 183 to 204, 211 to 227, and 250 to 271 of SEQ ID NO: 55 arethe transmembrane domains of human INTERCEPT 296 (SEQ ID NOs: 59 through63, respectively). As described elsewhere herein, relatively hydrophilicregions are generally located at or near the surface of a protein, andare more frequently effective immunogenic epitopes than are relativelyhydrophobic regions. For example, the region of human INTERCEPT 296protein from about amino acid residue 120 to about amino acid residue140 appears to be located at or near the surface of the protein, whilethe region from about amino acid residue 95 to about amino acid residue110 appears not to be located at or near the surface.

[0310] The predicted molecular weight of INTERCEPT 296 protein withoutmodification and prior to cleavage of the signal sequence is about 37.8kilodaltons. The predicted molecular weight of the mature INTERCEPT 296protein without modification and after cleavage of the signal sequenceis about 30.2 kilodaltons.

[0311]FIGS. 7E and 7F depicts an alignment of the amino acid sequencesof human INTERCEPT 296 protein (SEQ ID NO: 55) and Caenorhabditiselegans C06E1.3 related protein (SEQ ID NO: 399). In this alignment(pam120.mat scoring matrix, gap penalties −12/−4), the amino acidsequences of the proteins are 26.8% identical. The C. elegans proteinhas five predicted transmembrane domains.

[0312] Biological Function of INTERCEPT 296 Proteins, Nucleic Acids, andModulators Thereof

[0313] The cDNA encoding INTERCEPT 296 protein was obtained from a humanesophagus cDNA library, and INTERCEPT 296 is expressed in lung tissue.The INTERCEPT 296-related proteins and nucleic acids of the inventionare therefore useful for prevention, detection, and treatment ofdisorders of the human lung and esophagus. Such disorders include, forexample, various cancers, bronchitis, cystic fibrosis, respiratoryinfections (e.g., influenza, bronchiolitis, pneumonia, andtuberculosis), asthma, emphysema, chronic bronchitis, bronchiectasis,pulmonary edema, pleural effusion, pulmonary embolus, adult and infantrespiratory distress syndromes, heartburn, and gastric reflux esophagealdisease.

[0314] Tables A and B summarize sequence data corresponding to the humanproteins herein designated TANGO 202, TANGO 234, TANGO 265, TANGO 273,TANGO 286, TANGO 294, and INTERCEPT 296. TABLE A Protein SEQ ID NOsDepicted in ATCC ® Designation cDNA ORF Protein Figure # Accession #TANGO 202 1 2 3 1 207219 TANGO 234 9 10 11 2 207184 TANGO 265 17 18 19 3207228 TANGO 273 25 26 27 4 207185 TANGO 286 33 34 35 5 207220 TANGO 29445 46 47 6 207220 INTERCEPT 53 54 55 7 207220 296

[0315] TABLE B Amino Acid Residues SEQ ID NOs ExtracellularTransmembrane Cytoplasmic Protein Desig. Signal Sequence Mature ProteinDomain(s) Domain(s) Domain(s) TANGO 202 1 to 19  4 20 to 475  5 20 to392  6 393 to 415  7 416 to 475  8 (variant) (1 to 19)  (4) (20 to 475) (5) (20 to 475)  (5) (N/A) (N/A) TANGO 234 1 to 40 12  41 to 1453 13 41 to 1359 14 1360 to 1383 15 1384 to 1453 16 TANGO 265 1 to 31 20 32to 761 21 32 to 683 22 684 to 704 23 705 to 761 24 TANGO 273 1 to 22 2823 to 172 29 23 to 60  30 61 to 81 31  82 to 172 32 TANGO 286 1 to 23 3624 to 455 37 24 to 455 37 N/A N/A TANGO 294 1 to 33 48 34 to 423 49 34to 254 50 255 to 279 51 280 to 423 52 (variant 1) (15 to 33)  (40) (34to 423) (49) (34 to 254) (50) (255 to 279) (51) (280 to 423) (52)<variant 2> <1 to 33> <48> <34 to 423> <49> <34 to 423> <49> <N/A> <N/A>{variant 3} {15 to 33}  {40} {34 to 423} {49} {34 to 423} {49} { N/A } {N/A } INTERCEPT N/A  1 to 343 55  1 to 23  56  24 to 42  59  43 to 50 64 296 71 to 182 57  51 to 70  60 205 to 210 65 228 to 249  58 183 to204 61 272 to 343 66 211 to 227 62 250 to 271 63

[0316] Various aspects of the invention are described in further detailin the following subsections.

[0317] Isolated Nucleic Acid Molecules

[0318] One aspect of the invention pertains to isolated nucleic acidmolecules that encode a polypeptide of the invention or a biologicallyactive portion thereof, as well as nucleic acid molecules sufficient foruse as hybridization probes to identify nucleic acid molecules encodinga polypeptide of the invention and fragments of such nucleic acidmolecules suitable for use as PCR primers for the amplification ormutation of nucleic acid molecules. As used herein, the term “nucleicacid molecule” is intended to include DNA molecules (e.g., cDNA orgenomic DNA) and RNA molecules (e.g., mRNA) and analogs of the DNA orRNA generated using nucleotide analogs. The nucleic acid molecule can besingle-stranded or double-stranded, but preferably is double-strandedDNA.

[0319] An “isolated” nucleic acid molecule is one which is separatedfrom other nucleic acid molecules which are present in the naturalsource of the nucleic acid molecule. Preferably, an “isolated” nucleicacid molecule is free of sequences (preferably protein-encodingsequences) which naturally flank the nucleic acid (i.e., sequenceslocated at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA ofthe organism from which the nucleic acid is derived. For example, invarious embodiments, the isolated nucleic acid molecule can contain lessthan about 5 kB, 4 kB, 3 kB, 2 kB, 1 kB, 0.5 kB or 0.1 kB of nucleotidesequences which naturally flank the nucleic acid molecule in genomic DNAof the cell from which the nucleic acid is derived. Moreover, an“isolated” nucleic acid molecule, such as a cDNA molecule, can besubstantially free of other cellular material, or culture medium whenproduced by recombinant techniques, or substantially free of chemicalprecursors or other chemicals when chemically synthesized.

[0320] A nucleic acid molecule of the present invention, e.g., a nucleicacid molecule having the nucleotide sequence of all or a portion of anyof SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67,68, 72, and 73, or a complement thereof, or which has a nucleotidesequence comprising one of these sequences, can be isolated usingstandard molecular biology techniques and the sequence informationprovided herein. Using a nucleic acid comprising at least one of thesequences of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46,53, 54, 67, 68, 72, and 73 as a hybridization probe, nucleic acidmolecules of the invention can be isolated using standard hybridizationand cloning techniques (e.g., as described in Sambrook et al., eds.,Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring HarborLaboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor,N.Y., 1989).

[0321] A nucleic acid molecule of the invention can be amplified usingcDNA, mRNA or genomic DNA as a template and appropriate oligonucleotideprimers according to standard PCR amplification techniques. The nucleicacid so amplified can be cloned into an appropriate vector andcharacterized by DNA sequence analysis. Furthermore, oligonucleotidescorresponding to all or a portion of a nucleic acid molecule of theinvention can be prepared by standard synthetic techniques, e.g., usingan automated DNA synthesizer.

[0322] In another preferred embodiment, an isolated nucleic acidmolecule of the invention comprises a nucleic acid molecule which is acomplement of the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10,17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or a portionthereof. A nucleic acid molecule which is complementary to a givennucleotide sequence is one which is sufficiently complementary to thegiven nucleotide sequence that it can hybridize to the given nucleotidesequence thereby forming a stable duplex.

[0323] Moreover, a nucleic acid molecule of the invention can compriseonly a portion of a nucleic acid sequence encoding a full lengthpolypeptide of the invention for example, a fragment which can be usedas a probe or primer or a fragment encoding a biologically activeportion of a polypeptide of the invention. The nucleotide sequencedetermined from the cloning one gene allows for the generation of probesand primers designed for use in identifying and/or cloning homologs inother cell types, e.g., from other tissues, as well as homologs fromother mammals. The probe/primer typically comprises substantiallypurified oligonucleotide. The oligonucleotide typically comprises aregion of nucleotide sequence that hybridizes under stringent conditionsto at least about 15, preferably about 25, more preferably about 50, 75,100, 125, 150, 175, 200, 250, 300, 350, or 400 or more consecutivenucleotides of the sense or anti-sense sequence of any of SEQ ID NOs: 1,2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, orof a naturally occurring mutant of any of SEQ ID NOs: 1, 2, 9, 10, 17,18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73.

[0324] Probes based on the sequence of a nucleic acid molecule of theinvention can be used to detect transcripts or genomic sequencesencoding the same protein molecule encoded by a selected nucleic acidmolecule. The probe comprises a label group attached thereto, e.g., aradioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor.Such probes can be used as part of a diagnostic test kit for identifyingcells or tissues which mis-express the protein, such as by measuringlevels of a nucleic acid molecule encoding the protein in a sample ofcells from a subject, e.g., detecting mRNA levels or determining whethera gene encoding the protein has been mutated or deleted.

[0325] A nucleic acid fragment encoding a biologically active portion ofa polypeptide of the invention can be prepared by isolating a portion ofany of SEQ ID NOs: 2, 10, 18, 26, 34, 46, 54, 68, and 73, expressing theencoded portion of the polypeptide protein (e.g., by recombinantexpression in vitro), and assessing the activity of the encoded portionof the polypeptide.

[0326] The invention further encompasses nucleic acid molecules thatdiffer from the nucleotide sequence of any of SEQ ID NOs: 1, 2, 9, 10,17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 due todegeneracy of the genetic code and thus encode the same protein as thatencoded by the nucleotide sequence of any of SEQ ID NOs: 2, 10, 18, 26,34, 46, 54, 68,and 73.

[0327] In addition to the nucleotide sequences of SEQ ID NOs: 2, 10, 18,26, 34, 46, 54, 68, and 73, it will be appreciated by those skilled inthe art that DNA sequence polymorphisms that lead to changes in theamino acid sequence can exist within a population (e.g., the humanpopulation). Such genetic polymorphisms can exist among individualswithin a population due to natural allelic variation. An allele is oneof a group of genes which occur alternatively at a given genetic locus.

[0328] As used herein, the phrase “allelic variant” refers to anucleotide sequence which occurs at a given locus or to a polypeptideencoded by the nucleotide sequence. For example, chromosomal mapping hasbeen used to locate the gene encoding human TANGO 234 at chromosomallocation h12p13 (with synteny to mo6), between chromosomal markersWI-6980 and GATA8A09.43. Thus, human TANGO 234 allelic variants caninclude TANGO 234 nucleotide sequence polymorphisms (e.g., nucleotidesequences that vary from SEQ ID NO: 9) that map to this chromosomalregion. Similarly, chromosomal mapping has been used to locate the geneencoding human TANGO 265 protein on chromosome 1, between markers D1S305and D1S2635. Allelic variants of TANGO 265 occur at this chromosomallocation. Further by way of example, the gene encoding human TANGO 273protein has been located by chromosomal mapping on chromosome 7, betweenmarkers D7S2467 and D7S2552. Allelic variants of TANGO 273 occur at thischromosomal location.

[0329] As used herein, the terms “gene” and “recombinant gene” refer tonucleic acid molecules comprising an open reading frame encoding apolypeptide of the invention. Such natural allelic variations cantypically result in 1-5% variance in the nucleotide sequence of a givengene. Alternative alleles can be identified by sequencing the gene ofinterest in a number of different individuals. This can be readilycarried out by using hybridization probes to identify the same geneticlocus in a variety of individuals. Any and all such nucleotidevariations and resulting amino acid polymorphisms or variations that arethe result of natural allelic variation and that do not alter thefunctional activity are intended to be within the scope of theinvention.

[0330] Moreover, nucleic acid molecules encoding proteins of theinvention from other species (homologs), which have a nucleotidesequence which differs from that of the specific proteins describedherein are intended to be within the scope of the invention. Nucleicacid molecules corresponding to natural allelic variants and homologs ofa cDNA of the invention can be isolated based on their homology withnucleic acid molecules described herein, using the specific cDNAsdescribed herein, or a portion thereof, as a hybridization probeaccording to standard hybridization techniques under stringenthybridization conditions. For example, a cDNA encoding a soluble form ofa membrane-bound protein of the invention isolated based on itshybridization to a nucleic acid molecule encoding all or part of themembrane-bound form. Likewise, a cDNA encoding a membrane-bound form canbe isolated based on its hybridization to a nucleic acid moleculeencoding all or part of the soluble form.

[0331] Accordingly, in another embodiment, an isolated nucleic acidmolecule of the invention is at least 15 (25, 40, 60, 80, 100, 150, 200,250, 300, 350, 400, 450, 550, 650, 700, 800, 900, 1000, 1200, 1400,1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3500, 4000, 4500, or4928) nucleotides in length and hybridizes under stringent conditions tothe nucleic acid molecule comprising the nucleotide sequence, preferablythe coding sequence, of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73, or a complement thereof. Asused herein, the term “hybridizes under stringent conditions” isintended to describe conditions for hybridization and washing underwhich nucleotide sequences at least 60% (65%, 70%, preferably 75%)identical to each other typically remain hybridized to each other. Suchstringent conditions are known to those skilled in the art and can befound in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y.(1989), 6.3.1-6.3.6. A preferred, non-limiting example of stringenthybridization conditions are hybridization in 6× sodium chloride/sodiumcitrate (SSC) at about 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 50-65° C. Preferably, an isolated nucleic acid moleculeof the invention that hybridizes under stringent conditions with thesequence of any of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45,46, 53, 54, 67, 68, 72, and 73, or a complement thereof, corresponds toa naturally-occurring nucleic acid molecule. As used herein, a“naturally-occurring” nucleic acid molecule refers to an RNA or DNAmolecule having a nucleotide sequence that occurs in nature (e.g.,encodes a natural protein).

[0332] In addition to naturally-occurring allelic variants of a nucleicacid molecule of the invention sequence that can exist in thepopulation, the skilled artisan will further appreciate that changes canbe introduced by mutation thereby leading to changes in the amino acidsequence of the encoded protein, without altering the biologicalactivity of the protein. For example, one can make nucleotidesubstitutions leading to amino acid substitutions at “non-essential”amino acid residues. A “non-essential” amino acid residue is a residuethat can be altered from the wild-type sequence without altering thebiological activity, whereas an “essential” amino acid residue isrequired for biological activity. For example, amino acid residues thatare not conserved or only semi-conserved among homologs of variousspecies may be non-essential for activity and thus would be likelytargets for alteration. Alternatively, amino acid residues that areconserved among the homologs of various species (e.g., murine and human)may be essential for activity and thus would not be likely targets foralteration.

[0333] Accordingly, another aspect of the invention pertains to nucleicacid molecules encoding a polypeptide of the invention that containchanges in amino acid residues that are not essential for activity. Suchpolypeptides differ in amino acid sequence from the sequence of any ofSEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74,yet retain biological activity. In one embodiment, the isolated nucleicacid molecule includes a nucleotide sequence encoding a protein thatincludes an amino acid sequence that is at least about 40% identical,50%, 60%, 70%, 80%, 90%, 95%, or 98% identical to the amino acidsequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52,55-66, 69, and 74.

[0334] An isolated nucleic acid molecule encoding a variant protein canbe created by introducing one or more nucleotide substitutions,additions or deletions into the nucleotide sequence of any of SEQ IDNOs: 1, 2, 9, 10, 17, 18, 25, 26, 33, 34, 45, 46, 53, 54, 67, 68, 72,and 73, such that one or more amino acid residue substitutions,additions or deletions are introduced into the encoded protein.Mutations can be introduced by standard techniques, such assite-directed mutagenesis and PCR-mediated mutagenesis. Preferably,conservative amino acid substitutions are made at one or more predictednon-essential amino acid residues. A “conservative amino acidsubstitution” is one in which the amino acid residue is replaced with anamino acid residue having a similar side chain. Families of amino acidresidues having similar side chains have been defined in the art. Thesefamilies include amino acids with basic side chains (e.g., lysine,arginine, histidine), acidic side chains (e.g., aspartic acid, glutamicacid), uncharged polar side chains (e.g., glycine, asparagine,glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains(e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine,methionine, tryptophan), beta-branched side chains (e.g., threonine,valine, isoleucine) and aromatic side chains (e.g., tyrosine,phenylalanine, tryptophan, histidine). Alternatively, mutations can beintroduced randomly along all or part of the coding sequence, such as bysaturation mutagenesis, and the resultant mutants can be screened forbiological activity to identify mutants that retain activity. Followingmutagenesis, the encoded protein can be expressed recombinantly and theactivity of the protein can be determined.

[0335] In a preferred embodiment, a mutant polypeptide that is a variantof a polypeptide of the invention can be assayed for: (1) the ability toform protein:protein interactions with one or more polypeptides of theinvention (e.g., in a signaling pathway); (2) the ability to bind aligand of a polypeptide of the invention (e.g., another proteinidentified herein); (3) the ability to bind to an intracellular targetprotein of a polypeptide of the invention (e.g., a modulator orsubstrate of the polypeptide); or (4) the ability to modulate aphysiological activity of the protein, such as one of those disclosedherein (e.g., ability to modulate cell proliferation, cell migration,chemotaxis, or cellular differentiation).

[0336] The present invention encompasses antisense nucleic acidmolecules, i.e., molecules which are complementary to a sense nucleicacid encoding a polypeptide of the invention, e.g., complementary to thecoding strand of a double-stranded cDNA molecule or complementary to anmRNA sequence. Accordingly, an antisense nucleic acid can hydrogen bondto a sense nucleic acid. The antisense nucleic acid can be complementaryto an entire coding strand, or to only a portion thereof, e.g., all orpart of the protein coding region (or open reading frame). An antisensenucleic acid molecule can be antisense to all or part of a non-codingregion of the coding strand of a nucleotide sequence encoding apolypeptide of the invention. The non-coding regions (“5′ and 3′untranslated regions”) are the 5′ and 3′ sequences which flank thecoding region and are not translated into amino acids.

[0337] An antisense oligonucleotide can be, for example, about 5, 10,15, 20, 25, 30, 35, 40, 45, or 50 or more nucleotides in length. Anantisense nucleic acid of the invention can be constructed usingchemical synthesis and enzymatic ligation reactions using proceduresknown in the art. For example, an antisense nucleic acid (e.g., anantisense oligonucleotide) can be chemically synthesized using naturallyoccurring nucleotides or variously modified nucleotides designed toincrease the biological stability of the molecules or to increase thephysical stability of the duplex formed between the antisense and sensenucleic acids, e.g., phosphorothioate derivatives and acridinesubstituted nucleotides can be used. Examples of modified nucleotideswhich can be used to generate the antisense nucleic acid include5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil,hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl)uracil, 5-carboxymethylaminomethyl-2-thiouridine,5-carboxymethylaminomethyluracil, dihydrouracil,beta-D-galactosylqueosine, inosine, N₆-isopentenyladenine,1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine,2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine,7-methylguanine, 5-methylaminomethyluracil,5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine,5′-methoxycarboxymethyluracil, 5-methoxyuracil,2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v),wybutoxosine, pseudouracil, queosine, 2-thiocytosine,5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil,uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v),5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w,and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can beproduced biologically using an expression vector into which a nucleicacid has been sub-cloned in an antisense orientation (i.e., RNAtranscribed from the inserted nucleic acid will be of an antisenseorientation to a target nucleic acid of interest, described further inthe following subsection).

[0338] The antisense nucleic acid molecules of the invention aretypically administered to a subject or generated in situ such that theyhybridize with or bind to cellular mRNA and/or genomic DNA encoding aselected polypeptide of the invention to thereby inhibit expression,e.g., by inhibiting transcription and/or translation. The hybridizationcan be by conventional nucleotide complementarity to form a stableduplex, or, for example, in the case of an antisense nucleic acidmolecule which binds to DNA duplexes, through specific interactions inthe major groove of the double helix. An example of a route ofadministration of antisense nucleic acid molecules of the inventionincludes direct injection at a tissue site. Alternatively, antisensenucleic acid molecules can be modified to target selected cells and thenadministered systemically. For example, for systemic administration,antisense molecules can be modified such that they specifically bind toreceptors or antigens expressed on a selected cell surface, e.g., bylinking the antisense nucleic acid molecules to peptides or antibodieswhich bind to cell surface receptors or antigens. The antisense nucleicacid molecules can also be delivered to cells using the vectorsdescribed herein. To achieve sufficient intracellular concentrations ofthe antisense molecules, vector constructs in which the antisensenucleic acid molecule is placed under the control of a strong pol II orpol III promoter are preferred.

[0339] An antisense nucleic acid molecule of the invention can be analpha-anomeric nucleic acid molecule. An a-anomeric nucleic acidmolecule forms specific double-stranded hybrids with complementary RNAin which, contrary to the usual beta-units, the strands run parallel toeach other (Gaultier et al. (1987) Nucleic Acids Res. 15:6625-6641). Theantisense nucleic acid molecule can also comprise a2′-o-methylribonucleotide (Inoue et al. (1987) Nucleic Acids Res.15:6131-6148) or a chimeric RNA-DNA analogue (Inoue et al. (1987) FEBSLett. 215:327-330).

[0340] The invention also encompasses ribozymes. Ribozymes are catalyticRNA molecules with ribonuclease activity which are capable of cleaving asingle-stranded nucleic acid, such as an mRNA, to which they have acomplementary region. Thus, ribozymes (e.g., hammerhead ribozymes asdescribed in Haselhoff and Gerlach (1988) Nature 334:585-591) can beused to catalytically cleave mRNA transcripts to thereby inhibittranslation of the protein encoded by the mRNA. A ribozyme havingspecificity for a nucleic acid molecule encoding a polypeptide of theinvention can be designed based upon the nucleotide sequence of a cDNAdisclosed herein. For example, a derivative of a Tetrahymena L-19 IVSRNA can be constructed in which the nucleotide sequence of the activesite is complementary to the nucleotide sequence to be cleaved in a Cechet al. U.S. Pat. No. 4,987,071; and Cech et al. U.S. Pat. No. 5,116,742.Alternatively, an mRNA encoding a polypeptide of the invention can beused to select a catalytic RNA having a specific ribonuclease activityfrom a pool of RNA molecules. See, e.g., Bartel and Szostak (1993)Science 261:1411-1418.

[0341] The invention also encompasses nucleic acid molecules which formtriple helical structures. For example, expression of a polypeptide ofthe invention can be inhibited by targeting nucleotide sequencescomplementary to the regulatory region of the gene encoding thepolypeptide (e.g., the promoter and/or enhancer) to form triple helicalstructures that prevent transcription of the gene in target cells. Seegenerally Helene (1991) Anticancer Drug Des. 6(6):569-84; Helene (1992)Ann. N.Y. Acad. Sci. 660:27-36; and Maher (1992) Bioassays14(12):807-15.

[0342] In various embodiments, the nucleic acid molecules of theinvention can be modified at the base moiety, sugar moiety or phosphatebackbone to improve, e.g., the stability, hybridization, or solubilityof the molecule. For example, the deoxyribose phosphate backbone of thenucleic acids can be modified to generate peptide nucleic acids (seeHyrup et al. (1996) Bioorganic & Medicinal Chemistry 4(1): 5-23). Asused herein, the terms “peptide nucleic acids” or “PNAs” refer tonucleic acid mimics, e.g., DNA mimics, in which the deoxyribosephosphate backbone is replaced by a pseudopeptide backbone and only thefour natural nucleobases are retained. The neutral backbone of PNAs hasbeen shown to allow for specific hybridization to DNA and RNA underconditions of low ionic strength. The synthesis of PNA oligomers can beperformed using standard solid phase peptide synthesis protocols asdescribed in Hyrup et al. (1996), supra; Perry-O'Keefe et al. (1996)Proc. Natl. Acad. Sci. USA 93: 14670-675.

[0343] PNAs can be used in therapeutic and diagnostic applications. Forexample, PNAs can be used as antisense or anti-gene agents forsequence-specific modulation of gene expression by, e.g., inducingtranscription or translation arrest or inhibiting replication. PNAs canalso be used, e.g., in the analysis of single base pair mutations in agene by, e.g., PNA directed PCR clamping; as artificial restrictionenzymes when used in combination with other enzymes, e.g., S1 nucleases(Hyrup (1996), supra; or as probes or primers for DNA sequence andhybridization (Hyrup (1996), supra; Perry-O'Keefe et al. (1996) Proc.Natl. Acad. Sci. USA 93: 14670-675).

[0344] In another embodiment, PNAs can be modified, e.g., to enhancetheir stability or cellular uptake, by attaching lipophilic or otherhelper groups to PNA, by the formation of PNA-DNA chimeras, or by theuse of liposomes or other techniques of drug delivery known in the art.For example, PNA-DNA chimeras can be generated which can combine theadvantageous properties of PNA and DNA. Such chimeras allow DNArecognition enzymes, e.g., RNase H and DNA polymerases, to interact withthe DNA portion while the PNA portion would provide high bindingaffinity and specificity. PNA-DNA chimeras can be linked using linkersof appropriate lengths selected in terms of base stacking, number ofbonds between the nucleobases, and orientation (Hyrup (1996), supra).The synthesis of PNA-DNA chimeras can be performed as described in Hyrup(1996), supra, and Finn et al. (1996) Nucleic Acids Res. 24(17):3357-63.For example, a DNA chain can be synthesized on a solid support usingstandard phosphoramidite coupling chemistry and modified nucleosideanalogs. Compounds such as 5′-(4-methoxytrityl)amino-5′-deoxy-thymidinephosphoramidite can be used as a link between the PNA and the 5′ end ofDNA (Mag et al. (1989) Nucleic Acids Res. 17:5973-88). PNA monomers arethen coupled in a step-wise manner to produce a chimeric molecule with a5′ PNA segment and a 3′ DNA segment (Finn et al. (1996) Nucleic AcidsRes. 24(17):3357-63). Alternatively, chimeric molecules can besynthesized with a 5′ DNA segment and a 3′ PNA segment (Peterser et al.(1975) Bioorganic Med. Chem. Lett. 5:1119-11124).

[0345] In other embodiments, the oligonucleotide can include otherappended groups such as peptides (e.g., for targeting host cellreceptors in vivo), or agents facilitating transport across the cellmembrane (see, e.g., Letsinger et al. (1989) Proc. Natl. Acad. Sci. USA86:6553-6556; Lemaitre et al. (1987) Proc. Natl. Acad. Sci. USA84:648-652; PCT Publication No. WO 88/09810) or the blood-brain barrier(see, e.g., PCT Publication No. WO 89/10134). In addition,oligonucleotides can be modified with hybridization-triggered cleavageagents (see, e.g., Krol et al. (1988) Bio/Techniques 6:958-976) orintercalating agents (see, e.g., Zon (1988) Pharm. Res. 5:539-549). Tothis end, the oligonucleotide can be conjugated to another molecule,e.g., a peptide, hybridization triggered cross-linking agent, transportagent, hybridization-triggered cleavage agent, etc.

[0346] Isolated Proteins and Antibodies

[0347] One aspect of the invention pertains to isolated proteins, andbiologically active portions thereof, as well as polypeptide fragmentssuitable for use as immunogens to raise antibodies directed against apolypeptide of the invention. In one embodiment, the native polypeptidecan be isolated from cells or tissue sources by an appropriatepurification scheme using standard protein purification techniques. Inanother embodiment, polypeptides of the invention are produced byrecombinant DNA techniques. Alternative to recombinant expression, apolypeptide of the invention can be synthesized chemically usingstandard peptide synthesis techniques.

[0348] An “isolated” or “purified” protein or biologically activeportion thereof is substantially free of cellular material or othercontaminating proteins from the cell or tissue source from which theprotein is derived, or substantially free of chemical precursors orother chemicals when chemically synthesized. The language “substantiallyfree of cellular material” includes preparations of protein in which theprotein is separated from cellular components of the cells from which itis isolated or recombinantly produced. Thus, protein that issubstantially free of cellular material includes preparations of proteinhaving less than about 30%, 20%, 10%, or 5% (by dry weight) ofheterologous protein (also referred to herein as a “contaminatingprotein”). When the protein or biologically active portion thereof isrecombinantly produced, it is also preferably substantially free ofculture medium, i.e., culture medium represents less than about 20%,10%, or 5% of the volume of the protein preparation. When the protein isproduced by chemical synthesis, it is preferably substantially free ofchemical precursors or other chemicals, i.e., it is separated fromchemical precursors or other chemicals which are involved in thesynthesis of the protein. Accordingly such preparations of the proteinhave less than about 30%, 20%, 10%, 5% (by dry weight) of chemicalprecursors or compounds other than the polypeptide of interest.

[0349] Biologically active portions of a polypeptide of the inventioninclude polypeptides comprising amino acid sequences sufficientlyidentical to or derived from the amino acid sequence of the protein(e.g., the amino acid sequence shown in any of SEQ ID NOs: 3-8, 11-16,19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74), which include feweramino acids than the full length protein, and exhibit at least oneactivity of the corresponding full-length protein. Typically,biologically active portions comprise a domain or motif with at leastone activity of the corresponding protein. A biologically active portionof a protein of the invention can be a polypeptide which is, forexample, 10, 25, 50, 100 or more amino acids in length. Moreover, otherbiologically active portions, in which other regions of the protein aredeleted, can be prepared by recombinant techniques and evaluated for oneor more of the functional activities of the native form of a polypeptideof the invention.

[0350] Preferred polypeptides have the amino acid sequence of any of SEQID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74. Otheruseful proteins are substantially identical (e.g., at least about 40%,preferably 50%, 60%, 70%, 80%, 90%, 95%, or 99%) to any of SEQ ID NOs:3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69, and 74 and retain thefunctional activity of the protein of the correspondingnaturally-occurring protein yet differ in amino acid sequence due tonatural allelic variation or mutagenesis.

[0351] To determine the percent identity of two amino acid sequences orof two nucleic acids, the sequences are aligned for optimal comparisonpurposes (e.g., gaps can be introduced in the sequence of a first aminoacid or nucleic acid sequence for optimal alignment with a second aminoor nucleic acid sequence). The amino acid residues or nucleotides atcorresponding amino acid positions or nucleotide positions are thencompared. When a position in the first sequence is occupied by the sameamino acid residue or nucleotide as the corresponding position in thesecond sequence, then the molecules are identical at that position. Thepercent identity between the two sequences is a function of the numberof identical positions shared by the sequences (i.e., % identity=# ofidentical positions/total # of positions (e.g., overlappingpositions)×100). In one embodiment the two sequences are the samelength.

[0352] The determination of percent identity between two sequences canbe accomplished using a mathematical algorithm. A preferred,non-limiting example of a mathematical algorithm utilized for thecomparison of two sequences is the algorithm of Karlin and Altschul(1990) Proc. Natl. Acad. Sci. USA 87:2264-2268, modified as in Karlinand Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877. Such analgorithm is incorporated into the NBLAST and XBLAST programs ofAltschul, et al. (1990) J. Mol. Biol. 215:403-410. BLAST nucleotidesearches can be performed with the NBLAST program, score=100,wordlength=12 to obtain nucleotide sequences homologous to a nucleicacid molecules of the invention. BLAST protein searches can be performedwith the XBLAST program, score=50, wordlength=3 to obtain amino acidsequences homologous to a protein molecules of the invention. To obtaingapped alignments for comparison purposes, Gapped BLAST can be utilizedas described in Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402.Alternatively, PSI-Blast can be used to perform an iterated search whichdetects distant relationships between molecules. Id. When utilizingBLAST, Gapped BLAST, and PSI-Blast programs, the default parameters ofthe respective programs (e.g., XBLAST and NBLAST) can be used. Seehttp://www.ncbi.nlm.nih.gov. Another preferred, non-limiting example ofa mathematical algorithm utilized for the comparison of sequences is thealgorithm of Myers and Miller, (1988) CABIOS 4:11-17. Such an algorithmis incorporated into the ALIGN program (version 2.0) which is part ofthe GCG sequence alignment software package. When utilizing the ALIGNprogram for comparing amino acid sequences, a PAM120 weight residuetable, a gap length penalty of 12, and a gap penalty of 4 can be used.

[0353] The percent identity between two sequences can be determinedusing techniques similar to those described above, with or withoutallowing gaps. In calculating percent identity, only exact matches arecounted.

[0354] The invention also provides chimeric or fusion proteins. As usedherein, a “chimeric protein” or “fusion protein” comprises all or part(preferably biologically active) of a polypeptide of the inventionoperably linked to a heterologous polypeptide (i.e., a polypeptide otherthan the same polypeptide of the invention). Within the fusion protein,the term “operably linked” is intended to indicate that the polypeptideof the invention and the heterologous polypeptide are fused in-frame toeach other. The heterologous polypeptide can be fused to theamino-terminus or the carboxyl-terminus of the polypeptide of theinvention.

[0355] One useful fusion protein is a GST fusion protein in which thepolypeptide of the invention is fused to the carboxyl terminus of GSTsequences. Such fusion proteins can facilitate the purification of arecombinant polypeptide of the invention.

[0356] In another embodiment, the fusion protein contains a heterologoussignal sequence at its amino terminus. For example, the native signalsequence of a polypeptide of the invention can be removed and replacedwith a signal sequence from another protein. For example, the gp67secretory sequence of the baculovirus envelope protein can be used as aheterologous signal sequence (Current Protocols in Molecular Biology,Ausubel et al., eds., John Wiley & Sons, 1992). Other examples ofeukaryotic heterologous signal sequences include the secretory sequencesof melittin and human placental alkaline phosphatase (Stratagene; LaJolla, Calif.). In yet another example, useful prokaryotic heterologoussignal sequences include the phoA secretory signal (Sambrook et al.,supra) and the protein A secretory signal (Pharmacia Biotech;Piscataway, N.J.).

[0357] In yet another embodiment, the fusion protein is animmunoglobulin fusion protein in which all or part of a polypeptide ofthe invention is fused to sequences derived from a member of theimmunoglobulin protein family. The immunoglobulin fusion proteins of theinvention can be incorporated into pharmaceutical compositions andadministered to a subject to inhibit an interaction between a ligand(soluble or membrane-bound) and a protein on the surface of a cell(receptor), to thereby suppress signal transduction in vivo. Theimmunoglobulin fusion protein can be used to affect the bioavailabilityof a cognate ligand of a polypeptide of the invention. Inhibition ofligand/receptor interaction can be useful therapeutically, both fortreating proliferative and differentiative disorders and for modulating(e.g., promoting or inhibiting) cell survival. Moreover, theimmunoglobulin fusion proteins of the invention can be used asimmunogens to produce antibodies directed against a polypeptide of theinvention in a subject, to purify ligands and in screening assays toidentify molecules which inhibit the interaction of receptors withligands.

[0358] Chimeric and fusion proteins of the invention can be produced bystandard recombinant DNA techniques. In another embodiment, the fusiongene can be synthesized by conventional techniques including automatedDNA synthesizers. Alternatively, PCR amplification of gene fragments canbe carried out using anchor primers which give rise to complementaryoverhangs between two consecutive gene fragments which can subsequentlybe annealed and re-amplified to generate a chimeric gene sequence (see,e.g., Ausubel et al., supra). Moreover, many expression vectors arecommercially available that already encode a fusion moiety (e.g., a GSTpolypeptide). A nucleic acid encoding a polypeptide of the invention canbe cloned into such an expression vector such that the fusion moiety islinked in-frame to the polypeptide of the invention.

[0359] A signal sequence of a polypeptide of the invention (e.g., thesignal sequence in one of SEQ ID NOs: 3, 4, 11, 12, 19, 20, 27, 28, 35,36, 47, 48, 69, and 74) can be used to facilitate secretion andisolation of the secreted protein or other proteins of interest. Signalsequences are typically characterized by a core of hydrophobic aminoacids which are generally cleaved from the mature protein duringsecretion in one or more cleavage events. Such signal peptides containprocessing sites that allow cleavage of the signal sequence from themature proteins as they pass through the secretory pathway. Thus, theinvention pertains to the described polypeptides having a signalsequence, as well as to the signal sequence itself and to thepolypeptide in the absence of the signal sequence (i.e., the cleavageproducts). In one embodiment, a nucleic acid sequence encoding a signalsequence of the invention can be operably linked in an expression vectorto a protein of interest, such as a protein which is ordinarily notsecreted or is otherwise difficult to isolate. The signal sequencedirects secretion of the protein, such as from a eukaryotic host intowhich the expression vector is transformed, and the signal sequence issubsequently or concurrently cleaved. The protein can then be readilypurified from the extracellular medium by art recognized methods.Alternatively, the signal sequence can be linked to the protein ofinterest using a sequence which facilitates purification, such as with aGST domain.

[0360] In another embodiment, the signal sequences of the presentinvention can be used to identify regulatory sequences, e.g., promoters,enhancers, repressors. Since signal sequences are the mostamino-terminal sequences of a peptide, it is expected that the nucleicacids which flank the signal sequence on its amino-terminal side will beregulatory sequences which affect transcription. Thus, a nucleotidesequence which encodes all or a portion of a signal sequence can be usedas a probe to identify and isolate signal sequences and their flankingregions, and these flanking regions can be studied to identifyregulatory elements therein.

[0361] The present invention also pertains to variants of thepolypeptides of the invention. Such variants have an altered amino acidsequence which can function as either agonists (mimetics) or asantagonists. Variants can be generated by mutagenesis, e.g., discretepoint mutation or truncation. An agonist can retain substantially thesame, or a subset, of the biological activities of the naturallyoccurring form of the protein. An antagonist of a protein can inhibitone or more of the activities of the naturally occurring form of theprotein by, for example, competitively binding to a downstream orupstream member of a cellular signaling cascade which includes theprotein of interest. Thus, specific biological effects can be elicitedby treatment with a variant of limited function. Treatment of a subjectwith a variant having a subset of the biological activities of thenaturally occurring form of the protein can have fewer side effects in asubject relative to treatment with the naturally occurring form of theprotein.

[0362] Variants of a protein of the invention which function as eitheragonists (mimetics) or as antagonists can be identified by screeningcombinatorial libraries of mutants, e.g., truncation mutants, of theprotein of the invention for agonist or antagonist activity. In oneembodiment, a variegated library of variants is generated bycombinatorial mutagenesis at the nucleic acid level and is encoded by avariegated gene library. A variegated library of variants can beproduced by, for example, enzymatically ligating a mixture of syntheticoligonucleotides into gene sequences such that a degenerate set ofpotential protein sequences is expressible as individual polypeptides,or alternatively, as a set of larger fusion proteins (e.g., for phagedisplay). There are a variety of methods which can be used to producelibraries of potential variants of the polypeptides of the inventionfrom a degenerate oligonucleotide sequence. Methods for synthesizingdegenerate oligonucleotides are known in the art (see, e.g., Narang(1983) Tetrahedron 39:3; Itakura et al. (1984) Annu. Rev. Biochem.53:323; Itakura et al. (1984) Science 198:1056; Ike et al. (1983)Nucleic Acid Res. 11:477).

[0363] In addition, libraries of fragments of the coding sequence of apolypeptide of the invention can be used to generate a variegatedpopulation of polypeptides for screening and subsequent selection ofvariants. For example, a library of coding sequence fragments can begenerated by treating a double stranded PCR fragment of the codingsequence of interest with a nuclease under conditions wherein nickingoccurs only about once per molecule, denaturing the double stranded DNA,re-naturing the DNA to form double stranded DNA which can includesense/antisense pairs from different nicked products, removing singlestranded portions from reformed duplexes by treatment with S1 nuclease,and ligating the resulting fragment library into an expression vector.By this method, an expression library can be derived which encodes aminoterminal and internal fragments of various sizes of the protein ofinterest.

[0364] Several techniques are known in the art for screening geneproducts of combinatorial libraries made by point mutations ortruncation, and for screening cDNA libraries for gene products having aselected property. The most widely used techniques, which are amenableto high through-put analysis, for screening large gene librariestypically include cloning the gene library into replicable expressionvectors, transforming appropriate cells with the resulting library ofvectors, and expressing the combinatorial genes under conditions inwhich detection of a desired activity facilitates isolation of thevector encoding the gene whose product was detected. Recursive ensemblemutagenesis (REM), a technique which enhances the frequency offunctional mutants in the libraries, can be used in combination with thescreening assays to identify variants of a protein of the invention(Arkin and Yourvan (1992) Proc. Natl. Acad. Sci. USA 89:7811-7815;Delgrave et al. (1993) Protein Engineering 6(3):327-331).

[0365] An isolated polypeptide of the invention, or a fragment thereof,can be used as an immunogen to generate antibodies using standardtechniques for polyclonal and monoclonal antibody preparation. Thefull-length polypeptide or protein can be used or, alternatively, theinvention provides antigenic peptide fragments for use as immunogens.The antigenic peptide of a protein of the invention comprises at least 8(preferably 10, 15, 20, or 30 or more) amino acid residues of the aminoacid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44,47-52, 55-66, 69, and 74, and encompasses an epitope of the protein suchthat an antibody raised against the peptide forms a specific immunecomplex with the protein.

[0366] Preferred epitopes encompassed by the antigenic peptide areregions that are located on the surface of the protein, e.g.,hydrophilic regions. FIGS. 1L, 1M, 2J, 3U, 4I, 4J, 5E, 6F, and 7D arehydrophobicity plots of the proteins of the invention. These plots orsimilar analyses can be used to identify hydrophilic regions.

[0367] An immunogen typically is used to prepare antibodies byimmunizing a suitable (i.e., immunocompetent) subject such as a rabbit,goat, mouse, or other mammal or vertebrate. An appropriate immunogenicpreparation can contain, for example, recombinantly-expressed orchemically-synthesized polypeptide. The preparation can further includean adjuvant, such as Freund's complete or incomplete adjuvant, or asimilar immunostimulatory agent.

[0368] Accordingly, another aspect of the invention pertains toantibodies directed against a polypeptide of the invention. The terms“antibody” and “antibody substance” as used interchangeably herein referto immunoglobulin molecules and immunologically active portions ofimmunoglobulin molecules, i.e., molecules that contain an antigenbinding site which specifically binds an antigen, such as a polypeptideof the invention (e.g., an epitope of a polypeptide of the invention). Amolecule which specifically binds to a given polypeptide of theinvention is a molecule which binds the polypeptide, but does notsubstantially bind other molecules in a sample, e.g., a biologicalsample, which naturally contains the polypeptide. Examples ofimmunologically active portions of immunoglobulin molecules includeF(ab) and F(ab′)₂ fragments which can be generated by treating theantibody with an enzyme such as pepsin. The invention providespolyclonal and monoclonal antibodies. The term “monoclonal antibody” or“monoclonal antibody composition”, as used herein, refers to apopulation of antibody molecules that contain only one species of anantigen binding site capable of immunoreacting with a particularepitope.

[0369] Polyclonal antibodies can be prepared as described above byimmunizing a suitable subject with a polypeptide of the invention as animmunogen. Preferred polyclonal antibody compositions are ones that havebeen selected for antibodies directed against (i.e., which bindspecifically with) one or more polypeptides of the invention.Particularly preferred polyclonal antibody preparations are ones thatcontain only antibodies directed against one or more polypeptides of theinvention. Particularly preferred immunogen compositions are those thatcontain no other human proteins such as, for example, immunogencompositions made using a non-human host cell for recombinant expressionof a polypeptide of the invention. In such a manner, the only humanepitope or epitopes recognized by the resulting antibody compositionsraised against this immunogen will be present as part of a polypeptideor polypeptides of the invention.

[0370] The antibody titer in the immunized subject can be monitored overtime by standard techniques, such as with an enzyme linked immunosorbentassay (ELISA) using immobilized polypeptide. If desired, the antibodymolecules can be harvested or isolated from the subject (e.g., from theblood or serum of the subject) and further purified by well-knowntechniques, such as protein A chromatography to obtain the IgG fraction.Alternatively, antibodies which bind specifically with a protein orpolypeptide of the invention can be selected or purified (e.g.,partially purified) using chromatographic methods, such as affinitychromatography. For example, a recombinantly expressed and purified (orpartially purified) protein of the invention can be produced asdescribed herein, and covalently or non-covalently coupled with a solidsupport such as, for example, a chromatography column. The column thusexhibits specific affinity for antibody substances which bindspecifically with the protein of the invention, and these antibodysubstances can be purified from a sample containing antibody substancesdirected against a large number of different epitopes, therebygenerating a substantially purified antibody substance composition,i.e., one that is substantially free of antibody substances which do notbind specifically with the protein. A substantially purified antibodycomposition, in this context, means an antibody sample that contains atmost only 30% (by dry weight) of contaminating antibodies directedagainst epitopes other than those on the desired protein or polypeptideof the invention, preferably at most 20%, more preferably at most 10%,most preferably at most 5% (by dry weight of the sample is contaminatingantibodies). A purified antibody composition means that at least 99% ofthe antibodies in the composition are directed against the desiredprotein or polypeptide of the invention.

[0371] At an appropriate time after immunization, e.g., when thespecific antibody titers are highest, antibody-producing cells can beobtained from the subject and used to prepare monoclonal antibodies bystandard techniques, such as the hybridoma technique originallydescribed by Kohler and Milstein (1975) Nature 256:495-497, the human Bcell hybridoma technique (Kozbor et al. (1983) Immunol. Today 4:72), theEBV-hybridoma technique (Cole et al. (1985), Monoclonal Antibodies andCancer Therapy, Alan R. Liss, Inc., pp. 77-96) or trioma techniques. Thetechnology for producing hybridomas is well known (see generally CurrentProtocols in Immunology (1994) Coligan et al. (eds.) John Wiley & Sons,Inc., New York, N.Y.). Hybridoma cells producing a monoclonal antibodyof the invention are detected by screening the hybridoma culturesupernatants for antibodies that bind the polypeptide of interest, e.g.,using a standard ELISA assay.

[0372] Alternative to preparing monoclonal antibody-secretinghybridomas, a monoclonal antibody directed against a polypeptide of theinvention can be identified and isolated by screening a recombinantcombinatorial immunoglobulin library (e.g., an antibody phage displaylibrary) with the polypeptide of interest. Kits for generating andscreening phage display libraries are commercially available (e.g., thePharmacia Recombinant Phage Antibody System, Catalog No. 27-9400-01; andthe Stratagene SURFZAP™ Phage Display Kit, Catalog No. 240612).Additionally, examples of methods and reagents particularly amenable foruse in generating and screening antibody display library can be foundin, for example, U.S. Pat. No. 5,223,409; PCT Publication No. WO92/18619; PCT Publication No. WO 91/17271; PCT Publication No. WO92/20791; PCT Publication No. WO 92/15679; PCT Publication No. WO93/01288; PCT Publication No. WO 92/01047; PCT Publication No. WO92/09690; PCT Publication No. WO 90/02809; Fuchs et al. (1991)Bio/Technology 9:1370-1372; Hay et al. (1992) Hum. Antibod. Hybridomas3:81-85; Huse et al. (1989) Science 246:1275-1281; Griffiths et al.(1993) EMBO J. 12:725-734.

[0373] Additionally, recombinant antibodies, such as chimeric andhumanized monoclonal antibodies, comprising both human and non-humanportions, which can be made using standard recombinant DNA techniques,are within the scope of the invention. A chimeric antibody is a moleculein which different portions of the antibody amino acid sequence arederived from different animal species, such as those having a variableregion derived from a murine monoclonal antibody and a constant regionderived from a human immunoglobulin. (See, e.g., Cabilly et al., U.S.Pat. No. 4,816,567; and Boss et al., U.S. Pat. No. 4,816,397). Humanizedantibodies are antibody molecules which are obtained from non-humanspecies, which have one or more complementarity-determining regions(CDRs) derived from the non-human species, and which have a frameworkregion derived from a human immunoglobulin molecule. (See, e.g., Queen,U.S. Pat. No. 5,585,089). Such chimeric and humanized monoclonalantibodies can be produced by recombinant DNA techniques known in theart, for example using methods described in PCT Publication No. WO87/02671; European Patent Application 184,187; European PatentApplication 171,496; European Patent Application 173,494; PCTPublication No. WO 86/01533; U.S. Pat. No. 4,816,567; European PatentApplication 125,023; Better et al. (1988) Science 240:1041-1043; Liu etal. (1987) Proc. Natl. Acad. Sci. USA 84:3439-3443; Liu et al. (1987) J.Immunol. 139:3521-3526; Sun et al. (1987) Proc. Natl. Acad. Sci. USA84:214-218; Nishimura et al. (1987) Canc. Res. 47:999-1005; Wood et al.(1985) Nature 314:446-449; and Shaw et al. (1988) J. Natl. Cancer Inst.80:1553-1559); Morrison (1985) Science 229:1202-1207; Oi et al. (1986)Bio/Techniques 4:214; U.S. Pat. No. 5,225,539; Jones et al. (1986)Nature 321:552-525; Verhoeyan et al. (1988) Science 239:1534; andBeidler et al. (1988) J. Immunol. 141:4053-4060.

[0374] Completely human antibodies are particularly desirable fortherapeutic treatment of human patients. Such antibodies can beproduced, for example, using transgenic mice which are incapable ofexpressing endogenous immunoglobulin heavy and light chains genes, butwhich can express human heavy and light chain genes The transgenic miceare immunized in the normal fashion with a selected antigen, e.g., allor a portion of a polypeptide of the invention. Monoclonal antibodiesdirected against the antigen can be obtained using conventionalhybridoma technology. The human immunoglobulin transgenes harbored bythe transgenic mice rearrange during B cell differentiation, andsubsequently undergo class switching and somatic mutation. Thus, usingsuch a technique, it is possible to produce therapeutically useful IgG,IgA and IgE antibodies. For an overview of this technology for producinghuman antibodies, see Lonberg and Huszar (1995, Int. Rev. Immunol.13:65-93). For a detailed discussion of this technology for producinghuman antibodies and human monoclonal antibodies and protocols forproducing such antibodies, see, e.g., U.S. Pat. Nos. 5,625,126;5,633,425; 5,569,825; 5,661,016; and 5,545,806. In addition, companiessuch as Abgenix, Inc. (Freemont, Calif.), can be engaged to providehuman antibodies directed against a selected antigen using technologysimilar to that described above.

[0375] Completely human antibodies which recognize a selected epitopecan be generated using a technique referred to as “guided selection.” Inthis approach a selected non-human monoclonal antibody, e.g., a murineantibody, is used to guide the selection of a completely human antibodyrecognizing the same epitope (Jespers et al. (1994) Bio/technology12:899-903).

[0376] An antibody directed against a polypeptide of the invention(e.g., monoclonal antibody) can be used to isolate the polypeptide bystandard techniques, such as affinity chromatography orimmunoprecipitation. Moreover, such an antibody can be used to detectthe protein (e.g., in a cellular lysate or cell supernatant) in order toevaluate the abundance and pattern of expression of the polypeptide. Theantibodies can also be used diagnostically to monitor protein levels intissue as part of a clinical testing procedure, e.g., to, for example,determine the efficacy of a given treatment regimen. Detection can befacilitated by coupling the antibody to a detectable substance. Examplesof detectable substances include various enzymes, prosthetic groups,fluorescent materials, luminescent materials, bioluminescent materials,and radioactive materials. Examples of suitable enzymes includehorseradish peroxidase, alkaline phosphatase, β-galactosidase, oracetylcholinesterase; examples of suitable prosthetic group complexesinclude streptavidin/biotin and avidin/biotin; examples of suitablefluorescent materials include umbelliferone, fluorescein, fluoresceinisothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansylchloride or phycoerythrin; an example of a luminescent material includesluminol; examples of bioluminescent materials include luciferase,luciferin, and aequorin, and examples of suitable radioactive materialinclude ¹²⁵I, ¹³¹I, ³⁵S or ³H.

[0377] Further, an antibody substance can be conjugated with atherapeutic moiety such as a cytotoxin, a therapeutic agent, or aradioactive metal ion. Cytotoxins and cytotoxic agents include any agentthat is detrimental to cells. Examples include taxol, cytochalasin B,gramicidin D, ethidium bromide, emetine, mitomycin, etoposide,tenoposide, vincristine, vinblastine, colchicin, doxorubicin,daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin,actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine,tetracaine, lidocaine, propranolol, puromycin, and analogs or homologsof these compounds. Therapeutic agents include, antimetabolites (e.g.,methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine,5-fluorouracil, and decarbazine), alkylating agents (e.g.,mechlorethamine, thioepa chlorambucil, melphalan, carmustine {BSNU},lomustine {CCNU}, cyclothosphamide, busulfan, dibromomannitol,streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP)cisplatin), anthracyclines (e.g., daunorubicin {formerly daunomycin} anddoxorubicin), antibiotics (e.g., dactinomycin {formerly actinomycin},bleomycin, mithramycin, and anthramycin {AMC}), and anti-mitotic agents(e.g., vincristine and vinblastine).

[0378] The conjugates of the invention can be used to modify abiological response; the drug moiety is not to be construed as limitedto classical chemical therapeutic agents. For example, the drug moietycan be a protein or polypeptide which exhibits a desired biologicalactivity. Such proteins include, for example, toxins such as abrin,ricin A, pseudomonas exotoxin, or diphtheria toxin; proteins such astumor necrosis factor, alpha-interferon, beta-interferon, nerve growthfactor, platelet derived growth factor, tissue plasminogen activator;and biological response modifiers such as lymphokines, interleukin-1(IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), granulocytemacrophage colony stimulating factor (GM-CSF), granulocyte colonystimulating factor (G-CSF), and other growth factors.

[0379] Techniques for conjugating a therapeutic moiety with an antibodysubstance are well known (see, e.g., Arnon et al., “MonoclonalAntibodies For Immunotargeting Of Drugs In Cancer Therapy”, inMonoclonal Antibodies and Cancer Therapy, Reisfeld et al., eds., pp.243-256, Alan R. Liss, Inc., 1985; Hellstrom et al., “Antibodies ForDrug Delivery”, in Controlled Drug Delivery, 2nd Ed., Robinson et al.,eds., pp. 623-653, Marcel Dekker, Inc., 1987; Thorpe, “Antibody CarriersOf Cytotoxic Agents In Cancer Therapy: A Review”, in MonoclonalAntibodies '84: Biological and Clinical Applications, Pinchera et al.,eds., pp. 475-506, 1985; “Analysis, Results, And Future Prospective OfThe Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, inMonoclonal Antibodies for Cancer Detection and Therapy, Baldwin et al.,eds., pp. 303-316, Academic Press, 1985; and Thorpe et al., “ThePreparation And Cytotoxic Properties Of Antibody-Toxin Conjugates”,Immunol. Rev. 62:119-58, 1982). Alternatively, an antibody can beconjugated with a second antibody to form an antibody heteroconjugate asdescribed by Segal in U.S. Pat. No. 4,676,980.

[0380] Accordingly, in one aspect, the invention provides substantiallypurified antibodies or fragment thereof, and non-human antibodies orfragments thereof, which antibodies or fragments specifically bind witha polypeptide having an amino acid sequence which comprises a sequenceselected from the group consisting of:

[0381] (i) SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66,69, and 74;

[0382] (ii) the amino acid sequence encoded by a cDNA of a clonedeposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and207221;

[0383] (iii) at least 15 amino acid residues of the amino acid sequenceof any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69,and 74;

[0384] (iv) an amino acid sequence which is at least 95% identical tothe amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32,35-44, 47-52, 55-66, 69, and 74, wherein the percent identity isdetermined using the ALIGN program of the GCG software package with aPAM120 weight residue table, a gap length penalty of 12, and a gappenalty of 4; and

[0385] (v) an amino acid sequence which is encoded by a nucleic acidmolecule which hybridizes with a nucleic acid having a sequence selectedfrom the group consisting of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 under conditions ofhybridization of 6× SSC (standard saline citrate) at 45° C. and washingin 0.2× SSC, 0.1% SDS at 65° C.

[0386] In another aspect, the invention provides non-human antibodies orfragments thereof, which antibodies or fragments specifically bind witha polypeptide having an amino acid sequence which comprises a sequenceselected from the group consisting of:

[0387] (i) SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66,69, and 74;

[0388] (ii) the amino acid sequence encoded by a cDNA of a clonedeposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and207221;

[0389] (iii) at least 15 amino acid residues of the amino acid sequenceof any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69,and 74;

[0390] (iv) an amino acid sequence which is at least 95% identical tothe amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32,35-44, 47-52, 55-66, 69, and 74, wherein the percent identity isdetermined using the ALIGN program of the GCG software package with aPAM120 weight residue table, a gap length penalty of 12, and a gappenalty of 4; and

[0391] (v) an amino acid sequence which is encoded by a nucleic acidmolecule which hybridizes with a nucleic acid having a sequence selectedfrom the group consisting of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 under conditions ofhybridization of 6× SSC (standard saline citrate) at 45° C. and washingin 0.2× SSC, 0.1% SDS at 65° C. Such non-human antibodies can be goat,mouse, sheep, horse, chicken, rabbit, or rat antibodies. Alternatively,the non-human antibodies of the invention can be chimeric and/orhumanized antibodies. In addition, the non-human antibodies of theinvention can be polyclonal antibodies or monoclonal antibodies.

[0392] In still a further aspect, the invention provides monoclonalantibodies or fragments thereof, which antibodies or fragmentsspecifically bind with a polypeptide having an amino acid sequence whichcomprises a sequence selected from the group consisting of:

[0393] (i) SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66,69, and 74;

[0394] (ii) the amino acid sequence encoded by a cDNA of a clonedeposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and207221;

[0395] (iii) at least 15 amino acid residues of the amino acid sequenceof any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69,and 74;

[0396] (iv) an amino acid sequence which is at least 95% identical tothe amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32,35-44, 47-52, 55-66, 69, and 74, wherein the percent identity isdetermined using the ALIGN program of the GCG software package with aPAM120 weight residue table, a gap length penalty of 12, and a gappenalty of 4; and

[0397] (v) an amino acid sequence which is encoded by a nucleic acidmolecule which hybridizes with a nucleic acid having a sequence selectedfrom the group consisting of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 under conditions ofhybridization of 6× SSC (standard saline citrate) at 45° C. and washingin 0.2× SSC, 0.1% SDS at 65° C. The monoclonal antibodies can be human,humanized, chimeric and/or non-human antibodies.

[0398] The substantially purified antibodies or fragments thereof canspecifically bind with a signal peptide, a secreted sequence, anextracellular domain, a transmembrane or a cytoplasmic domaincytoplasmic membrane of a polypeptide of the invention. In aparticularly preferred embodiment, the substantially purified antibodiesor fragments thereof, the non-human antibodies or fragments thereofand/or the monoclonal antibodies or fragments thereof, of the inventionspecifically bind with a secreted sequence or with an extracellulardomain of one of TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286,TANGO 294, and INTERCEPT 296. Preferably, the extracellular domain withwhich the antibody substance binds has an amino acid sequence selectedfrom the group consisting of SEQ ID NOs: 5, 6, 14, 22, 30, 37, 49, 50,and 56-58.

[0399] Any of the antibody substances of the invention can be conjugatedwith a therapeutic moiety or to a detectable substance. Non-limitingexamples of detectable substances that can be conjugated with theantibody substances of the invention include an enzyme, a prostheticgroup, a fluorescent material (i.e., a fluorophore), a luminescentmaterial, a bioluminescent material, and a radioactive material (e.g., aradionuclide or a substituent comprising a radionuclide).

[0400] The invention also provides a kit containing an antibodysubstance of the invention conjugated with a detectable substance, andinstructions for use. Still another aspect of the invention is apharmaceutical composition comprising an antibody substance of theinvention and a pharmaceutically acceptable carrier. In preferredembodiments, the pharmaceutical composition contains an antibodysubstance of the invention, a therapeutic moiety (preferably conjugatedwith the antibody substance), and a pharmaceutically acceptable carrier.

[0401] Still another aspect of the invention is a method of making anantibody that specifically recognizes one of TANGO 202, TANGO 234, TANGO265, TANGO 273, TANGO 286, TANGO 294, and INTERCEPT 296. This methodcomprises immunizing a vertebrate (e.g., a mammal such as a rabbit,goat, or pig) with a polypeptide. The polypeptide used as an immunogenhas an amino acid sequence that comprises a sequence selected from thegroup consisting of:

[0402] (i) SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66,69, and 74;

[0403] (ii) the amino acid sequence encoded by a cDNA of a clonedeposited as one of ATCC® 207219, 207184, 207228, 207185, 207220, and207221;

[0404] (iii) at least 15 amino acid residues of the amino acid sequenceof any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32, 35-44, 47-52, 55-66, 69,and 74;

[0405] (iv) an amino acid sequence which is at least 95% identical tothe amino acid sequence of any of SEQ ID NOs: 3-8, 11-16, 19-24, 27-32,35-44, 47-52, 55-66, 69, and 74, wherein the percent identity isdetermined using the ALIGN program of the GCG software package with aPAM120 weight residue table, a gap length penalty of 12, and a gappenalty of 4; and

[0406] (v) an amino acid sequence which is encoded by a nucleic acidmolecule which hybridizes with a nucleic acid having a sequence selectedfrom the group consisting of SEQ ID NOs: 1, 2, 9, 10, 17, 18, 25, 26,33, 34, 45, 46, 53, 54, 67, 68, 72, and 73 under conditions ofhybridization of 6× SSC (standard saline citrate) at 45° C. and washingin 0.2× SSC, 0.1% SDS at 65° C.

[0407] After immunization, a sample is collected from the vertebratethat contains an antibody that specifically recognizes the polypeptidewith which the vertebrate was immunized. Preferably, the polypeptide isrecombinantly produced using a non-human host cell. Optionally, anantibody substance can be further purified from the sample usingtechniques well known to those of skill in the art. The method canfurther comprise making a monoclonal antibody-producing cell from a cellof the vertebrate. Optionally, antibodies can be collected from theantibody-producing cell.

[0408] Recombinant Expression Vectors and Host Cells

[0409] Another aspect of the invention pertains to vectors, preferablyexpression vectors, containing a nucleic acid encoding a polypeptide ofthe invention (or a portion thereof). As used herein, the term “vector”refers to a nucleic acid molecule capable of transporting anothernucleic acid to which it has been linked. One type of vector is a“plasmid”, which refers to a circular double stranded DNA loop intowhich additional DNA segments can be ligated. Another type of vector isa viral vector, wherein additional DNA segments can be ligated into theviral genome. Certain vectors are capable of autonomous replication in ahost cell into which they are introduced (e.g., bacterial vectors havinga bacterial origin of replication and episomal mammalian vectors). Othervectors (e.g., non-episomal mammalian vectors) are integrated into thegenome of a host cell upon introduction into the host cell, and therebyare replicated along with the host genome. Moreover, certain vectors,expression vectors, are capable of directing the expression of genes towhich they are operably linked. In general, expression vectors ofutility in recombinant DNA techniques are often in the form of plasmids(vectors). However, the invention is intended to include such otherforms of expression vectors, such as viral vectors (e.g., replicationdefective retroviruses, adenoviruses and adeno-associated viruses),which serve equivalent functions.

[0410] The recombinant expression vectors of the invention comprise anucleic acid of the invention in a form suitable for expression of thenucleic acid in a host cell. This means that the recombinant expressionvectors include one or more regulatory sequences, selected on the basisof the host cells to be used for expression, which is operably linked tothe nucleic acid sequence to be expressed. Within a recombinantexpression vector, “operably linked” is intended to mean that thenucleotide sequence of interest is linked to the regulatory sequence(s)in a manner which allows for expression of the nucleotide sequence(e.g., in an in vitro transcription/translation system or in a host cellwhen the vector is introduced into the host cell). The term “regulatorysequence” is intended to include promoters, enhancers and otherexpression control elements (e.g., polyadenylation signals). Suchregulatory sequences are described, for example, in Goeddel, GeneExpression Technology: Methods in Enzymology 185, Academic Press, SanDiego, Calif. (1990). Regulatory sequences include those which directconstitutive expression of a nucleotide sequence in many types of hostcell and those which direct expression of the nucleotide sequence onlyin certain host cells (e.g., tissue-specific regulatory sequences). Itwill be appreciated by those skilled in the art that the design of theexpression vector can depend on such factors as the choice of the hostcell to be transformed, the level of expression of protein desired, andthe like. The expression vectors of the invention can be introduced intohost cells to thereby produce proteins or peptides, including fusionproteins or peptides, encoded by nucleic acids as described herein.

[0411] The recombinant expression vectors of the invention can bedesigned for expression of a polypeptide of the invention in prokaryotic(e.g., E. coli) or eukaryotic cells (e.g., insect cells (usingbaculovirus expression vectors), yeast cells or mammalian cells).Suitable host cells are discussed further in Goeddel, supra.Alternatively, the recombinant expression vector can be transcribed andtranslated in vitro, for example using T7 promoter regulatory sequencesand T7 polymerase.

[0412] Expression of proteins in prokaryotes is most often carried outin E. coli with vectors containing constitutive or inducible promotersdirecting the expression of either fusion or non-fusion proteins. Fusionvectors add a number of amino acids to a protein encoded therein,usually to the amino terminus of the recombinant protein. Such fusionvectors typically serve three purposes: 1) to increase expression ofrecombinant protein; 2) to increase the solubility of the recombinantprotein; and 3) to aid in the purification of the recombinant protein byacting as a ligand in affinity purification. Often, in fusion expressionvectors, a proteolytic cleavage site is introduced at the junction ofthe fusion moiety and the recombinant protein to enable separation ofthe recombinant protein from the fusion moiety subsequent topurification of the fusion protein. Such enzymes, and their cognaterecognition sequences, include Factor Xa, thrombin and enterokinase.Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc;Smith and Johnson (1988) Gene 67:31-40), pMAL (New England Biolabs,Beverly, Mass.) and pRIT5 (Phannacia, Piscataway, N.J.) which fuseglutathione S-transferase (GST), maltose E binding protein, or proteinA, respectively, to the target recombinant protein.

[0413] Examples of suitable inducible non-fusion E. coli expressionvectors include pTrc (Amann et al., (1988) Gene 69:301-315) and pET 1 Id(Studier et al., Gene Expression Technology: Methods in Enzymology 185,Academic Press, San Diego, Calif. (1990) 60-89). Target gene expressionfrom the pTrc vector relies on host RNA polymerase transcription from ahybrid trp-lac fusion promoter. Target gene expression from the pET 11dvector relies on transcription from a T7 gn10-lac fusion promotermediated by a co-expressed viral RNA polymerase (T7 gn1). This viralpolymerase is supplied by host strains BL21(DE3) or HMS174(DE3) from aresident lambda prophage harboring a T7 gn1 gene under thetranscriptional control of the lacUV 5 promoter.

[0414] One strategy to maximize recombinant protein expression in E.coli is to express the protein in a host bacteria with an impairedcapacity to proteolytically cleave the recombinant protein (Gottesman,Gene Expression Technology: Methods in Enzymology 185, Academic Press,San Diego, Calif. (1990) 119-128). Another strategy is to alter thenucleic acid sequence of the nucleic acid to be inserted into anexpression vector so that the individual codons for each amino acid arethose preferentially utilized in E. coli (Wada et al. (1992) NucleicAcids Res. 20:2111-2118). Such alteration of nucleic acid sequences ofthe invention can be carried out by standard DNA synthesis techniques.

[0415] In another embodiment, the expression vector is a yeastexpression vector. Examples of vectors for expression in yeast S.cerevisiae include pYepSec1 (Baldari et al. (1987) EMBO J. 6:229-234),pMFa (Kurjan and Herskowitz, (1982) Cell 30:933-943), pJRY88 (Schultz etal. (1987) Gene 54:113-123), pYES2 (Invitrogen Corporation, San Diego,Calif.), and pPicZ (Invitrogen Corp, San Diego, Calif.).

[0416] Alternatively, the expression vector is a baculovirus expressionvector. Baculovirus vectors available for expression of proteins incultured insect cells (e.g., Sf 9 cells) include the pAc series (Smithet al. (1983) Mol. Cell Biol. 3:2156-2165) and the pVL series (Lucklowand Summers (1989) Virology 170:31-39).

[0417] In yet another embodiment, a nucleic acid of the invention isexpressed in mammalian cells using a mammalian expression vector.Examples of mammalian expression vectors include pCDM8 (Seed (1987)Nature 329:840) and pMT2PC (Kaufman et al. (1987) EMBO J. 6:187-195).When used in mammalian cells, the expression vector's control functionsare often provided by viral regulatory elements. For example, commonlyused promoters are derived from polyoma, Adenovirus 2, cytomegalovirusand Simian Virus 40. For other suitable expression systems for bothprokaryotic and eukaryotic cells see chapters 16 and 17 of Sambrook etal., supra.

[0418] In another embodiment, the recombinant mammalian expressionvector is capable of directing expression of the nucleic acidpreferentially in a particular cell type (e.g., tissue-specificregulatory elements are used to express the nucleic acid).Tissue-specific regulatory elements are known in the art. Non-limitingexamples of suitable tissue-specific promoters include the albuminpromoter (liver-specific; Pinkert et al. (1987) Genes Dev. 1:268-277),lymphoid-specific promoters (Calame and Eaton (1988) Adv. Immunol.43:235-275), in particular promoters of T cell receptors (Winoto andBaltimore (1989) EMBO J. 8:729-733) and immunoglobulins (Banerji et al.(1983) Cell 33:729-740; Queen and Baltimore (1983) Cell 33:741-748),neuron-specific promoters (e.g., the neurofilament promoter; Byrne andRuddle (1989) Proc. Natl. Acad. Sci. USA 86:5473-5477),pancreas-specific promoters (Edlund et al. (1985) Science 230:912-916),and mammary gland-specific promoters (e.g., milk whey promoter; U.S.Pat. No. 4,873,316 and European Application Publication No. 264,166).Developmentally-regulated promoters are also encompassed, for examplethe murine hox promoters (Kessel and Gruss (1990) Science 249:374-379)and the α-fetoprotein promoter (Campes and Tilghman (1989) Genes Dev.3:537-546).

[0419] The invention further provides a recombinant expression vectorcomprising a DNA molecule of the invention cloned into the expressionvector in an antisense orientation. That is, the DNA molecule isoperably linked to a regulatory sequence in a manner which allows forexpression (by transcription of the DNA molecule) of an RNA moleculewhich is antisense to the mRNA encoding a polypeptide of the invention.Regulatory sequences operably linked to a nucleic acid cloned in theantisense orientation can be chosen which direct the continuousexpression of the antisense RNA molecule in a variety of cell types, forinstance viral promoters and/or enhancers, or regulatory sequences canbe chosen which direct constitutive, tissue specific or cell typespecific expression of antisense RNA. The antisense expression vectorcan be in the form of a recombinant plasmid, phagemid, or attenuatedvirus in which antisense nucleic acids are produced under the control ofa high efficiency regulatory region, the activity of which can bedetermined by the cell type into which the vector is introduced. For adiscussion of the regulation of gene expression using antisense genessee Weintraub et al. (Reviews—Trends in Genetics, Vol. 1(1) 1986).

[0420] Another aspect of the invention pertains to host cells into whicha recombinant expression vector of the invention has been introduced.The terms “host cell” and “recombinant host cell” are usedinterchangeably herein. It is understood that such terms refer not onlyto the particular subject cell but to the progeny or potential progenyof such a cell. Because certain modifications may occur in succeedinggenerations due to either mutation or environmental influences, suchprogeny may not, in fact, be identical to the parent cell, but are stillincluded within the scope of the term as used herein.

[0421] A host cell can be any prokaryotic (e.g., E. coli) or eukaryoticcell (e.g., insect cells, yeast or mammalian cells).

[0422] Vector DNA can be introduced into prokaryotic or eukaryotic cellsvia conventional transformation or transfection techniques. As usedherein, the terms “transformation” and “transfection” are intended torefer to a variety of art-recognized techniques for introducing foreignnucleic acid into a host cell, including calcium phosphate or calciumchloride co-precipitation, DEAE-dextran-mediated transfection,lipofection, or electroporation. Suitable methods for transforming ortransfecting host cells can be found in Sambrook, et al. (supra), andother laboratory manuals.

[0423] For stable transfection of mammalian cells, it is known that,depending upon the expression vector and transfection technique used,only a small fraction of cells may integrate the foreign DNA into theirgenome. In order to identify and select these integrants, a gene thatencodes a selectable marker (e.g., for resistance to antibiotics) isgenerally introduced into the host cells along with the gene ofinterest. Preferred selectable markers include those which conferresistance to drugs, such as G418, hygromycin and methotrexate. Cellsstably transfected with the introduced nucleic acid can be identified bydrug selection (e.g., cells that have incorporated the selectable markergene will survive, while the other cells die).

[0424] In another embodiment, the expression characteristics of anendogenous nucleic acid within a cell, cell line, or microorganism(e.g., a TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO294, or INTERCEPT 296 nucleic acid, as described herein) can be modifiedby inserting a heterologous DNA regulatory element (i.e., one that isheterologous with respect to the endogenous gene) into the genome of thecell, stable cell line, or cloned microorganism. The inserted regulatoryelement can be operatively linked with the endogenous gene (e.g., TANGO202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT296) and thereby control, modulate, or activate the endogenous gene. Forexample, an endogenous TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO286, TANGO 294, or INTERCEPT 296 gene which is normally“transcriptionally silent” (i.e., a TANGO 202, TANGO 234, TANGO 265,TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 gene which is normallynot expressed, or is normally expressed only at only a very low level)can be activated by inserting a regulatory element which is capable ofpromoting expression of the gene in the cell, cell line, ormicroorganism. Alternatively, a transcriptionally silent, endogenousTANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO 294, orINTERCEPT 296 gene can be activated by inserting a promiscuousregulatory element that works across cell types.

[0425] A heterologous regulatory element can be inserted into a stablecell line or cloned microorganism such that it is operatively linkedwith and activates expression of an endogenous TANGO 202, TANGO 234,TANGO 265, TANGO 273, TANGO 286, TANGO 294, or INTERCEPT 296 gene, usingtechniques, such as targeted homologous recombination, which are wellknown to those of skill in the art (described e.g., in Chappel, U.S.Pat. No. 5,272,071; PCT publication No. WO 91/06667, published May 16,1991).

[0426] A host cell of the invention, such as a prokaryotic or eukaryotichost cell in culture, can be used to produce a polypeptide of theinvention. Accordingly, the invention further provides methods forproducing a polypeptide of the invention using the host cells of theinvention. In one embodiment, the method comprises culturing the hostcell of invention (into which a recombinant expression vector encoding apolypeptide of the invention has been introduced) in a suitable mediumsuch that the polypeptide is produced. In another embodiment, the methodfurther comprises isolating the polypeptide from the medium or the hostcell.

[0427] The host cells of the invention can also be used to producenon-human transgenic animals. For example, in one embodiment, a hostcell of the invention is a fertilized oocyte or an embryonic stem cellinto which a sequences encoding a polypeptide of the invention have beenintroduced. Such host cells can then be used to create non-humantransgenic animals in which exogenous sequences encoding a polypeptideof the invention have been introduced into their genome or homologousrecombinant animals in which endogenous encoding a polypeptide of theinvention sequences have been altered. Such animals are useful forstudying the function and/or activity of the polypeptide and foridentifying and/or evaluating modulators of polypeptide activity. Asused herein, a “transgenic animal” is a non-human animal, preferably amammal, more preferably a rodent such as a rat or mouse, in which one ormore of the cells of the animal includes a transgene. Other examples oftransgenic animals include non-human primates, sheep, dogs, cows, goats,chickens, amphibians, etc. A transgene is exogenous DNA which isintegrated into the genome of a cell from which a transgenic animaldevelops and which remains in the genome of the mature animal, therebydirecting the expression of an encoded gene product in one or more celltypes or tissues of the transgenic animal. As used herein, an“homologous recombinant animal” is a non-human animal, preferably amammal, more preferably a mouse, in which an endogenous gene has beenaltered by homologous recombination between the endogenous gene and anexogenous DNA molecule introduced into a cell of the animal, e.g., anembryonic cell of the animal, prior to development of the animal.

[0428] A transgenic animal of the invention can be created byintroducing nucleic acid encoding a polypeptide of the invention (or ahomologue thereof) into the male pronuclei of a fertilized oocyte, e.g.,by microinjection, retroviral infection, and allowing the oocyte todevelop in a pseudopregnant female foster animal. Intronic sequences andpolyadenylation signals can also be included in the transgene toincrease the efficiency of expression of the transgene. Atissue-specific regulatory sequence(s) can be operably linked to thetransgene to direct expression of the polypeptide of the invention toparticular cells. Methods for generating transgenic animals via embryomanipulation and microinjection, particularly animals such as mice, havebecome conventional in the art and are described, for example, in U.S.Pat. Nos. 4,736,866 and 4,870,009, U.S. Pat. No. 4,873,191, in Hogan,Manipulating the Mouse Embryo, Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y., 1986, and in Wakayama et al., 1999, Proc. Natl.Acad. Sci. USA 96:14984-14989. Similar methods can be used to produceother transgenic animals. A transgenic founder animal can be identifiedbased upon the presence of the transgene in its genome and/or expressionof mRNA encoding the transgene in tissues or cells of the animals. Atransgenic founder animal can then be used to breed additional animalscarrying the transgene. Moreover, transgenic animals carrying thetransgene can further be bred to other transgenic animals carrying othertransgenes.

[0429] To create a homologous recombinant animal, a vector is preparedwhich contains at least a portion of a gene encoding a polypeptide ofthe invention into which a deletion, addition or substitution has beenintroduced to thereby alter, e.g., functionally disrupt, the gene. In apreferred embodiment, the vector is designed such that, upon homologousrecombination, the endogenous gene is functionally disrupted (i.e., nolonger encodes a functional protein; also referred to as a “knock out”vector). Alternatively, the vector can be designed such that, uponhomologous recombination, the endogenous gene is mutated or otherwisealtered but still encodes functional protein (e.g., the upstreamregulatory region can be altered to thereby alter the expression of theendogenous protein). In the homologous recombination vector, the alteredportion of the gene is flanked at its 5′ and 3′ ends by additionalnucleic acid of the gene to allow for homologous recombination to occurbetween the exogenous gene carried by the vector and an endogenous genein an embryonic stem cell. The additional flanking nucleic acidsequences are of sufficient length for successful homologousrecombination with the endogenous gene. Typically, several kilobases offlanking DNA (both at the 5′ and 3′ ends) are included in the vector(see, e.g., Thomas and Capecchi (1987) Cell 51:503 for a description ofhomologous recombination vectors). The vector is introduced into anembryonic stem cell line (e.g., by electroporation) and cells in whichthe introduced gene has homologously recombined with the endogenous geneare selected (see, e.g., Li et al. (1992) Cell 69:915). The selectedcells are then injected into a blastocyst of an animal (e.g., a mouse)to form aggregation chimeras (see, e.g., Bradley in Teratocarcinomas andEmbryonic Stem Cells: A Practical Approach, Robertson, ed. (IRL, Oxford,1987) pp. 113-152). A chimeric embryo can then be implanted into asuitable pseudopregnant female foster animal and the embryo brought toterm. Progeny harboring the homologously recombined DNA in their germcells can be used to breed animals in which all cells of the animalcontain the homologously recombined DNA by germline transmission of thetransgene. Methods for constructing homologous recombination vectors andhomologous recombinant animals are described further in Bradley (1991)Current Opinion in Bio/Technology 2:823-829 and in PCT Publication Nos.WO 90/11354, WO 91/01140, WO 92/0968, and WO 93/04169.

[0430] In another embodiment, transgenic non-human animals can beproduced which contain selected systems which allow for regulatedexpression of the transgene. One example of such a system is thecre/loxP recombinase system of bacteriophage P1. For a description ofthe cre/loxP recombinase system, see, e.g., Lakso et al. (1992) Proc.Natl. Acad. Sci. USA 89:6232-6236. Another example of a recombinasesystem is the FLP recombinase system of Saccharomyces cerevisiae(O'Gorman et al. (1991) Science 251:1351-1355. If a cre/loxP recombinasesystem is used to regulate expression of the transgene, animalscontaining transgenes encoding both the Cre recombinase and a selectedprotein are required. Such animals can be provided through theconstruction of “double” transgenic animals, e.g., by mating twotransgenic animals, one containing a transgene encoding a selectedprotein and the other containing a transgene encoding a recombinase.

[0431] Clones of the non-human transgenic animals described herein canalso be produced according to the methods described in Wilmut et al.(1997) Nature 385:810-813 and PCT Publication Nos. WO 97/07668 and WO97/07669.

[0432] Pharmaceutical Compositions

[0433] The nucleic acid molecules, polypeptides, and antibodies (alsoreferred to herein as “active compounds”) of the invention can beincorporated into pharmaceutical compositions suitable foradministration. Such compositions typically comprise the nucleic acidmolecule, protein, or antibody and a pharmaceutically acceptablecarrier. As used herein the language “pharmaceutically acceptablecarrier” is intended to include any and all solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents, and the like, compatible with pharmaceuticaladministration. The use of such media and agents for pharmaceuticallyactive substances is well known in the art. Except insofar as anyconventional media or agent is incompatible with the active compound,use thereof in the compositions is contemplated. Supplementary activecompounds can also be incorporated into the compositions.

[0434] The invention includes methods for preparing pharmaceuticalcompositions for modulating the expression or activity of a polypeptideor nucleic acid of the invention. Such methods comprise formulating apharmaceutically acceptable carrier with an agent which modulatesexpression or activity of a polypeptide or nucleic acid of theinvention. Such compositions can further include additional activeagents. Thus, the invention further includes methods for preparing apharmaceutical composition by formulating a pharmaceutically acceptablecarrier with an agent which modulates expression or activity of apolypeptide or nucleic acid of the invention and one or more additionalactive compounds.

[0435] The agent which modulates expression or activity can, forexample, be a small molecule other than a nucleic acid, polypeptide, orantibody of the invention. For example, such small molecules includepeptides, peptidomimetics, amino acids, amino acid analogs,polynucleotides, polynucleotide analogs, nucleotides, nucleotideanalogs, organic or inorganic compounds (i.e., including heteroorganicand organometallic compounds) having a molecular weight less than about10,000 grams per mole, organic or inorganic compounds having a molecularweight less than about 5,000 grams per mole, organic or inorganiccompounds having a molecular weight less than about 1,000 grams permole, organic or inorganic compounds having a molecular weight less thanabout 500 grams per mole, and salts, esters, and other pharmaceuticallyacceptable forms of such compounds.

[0436] It is understood that appropriate doses of small molecule agentsand protein or polypeptide agents depends upon a number of factorswithin the ken of the ordinarily skilled physician, veterinarian, orresearcher. The dose(s) of these agents will vary, for example,depending upon the identity, size, and condition of the subject orsample being treated, further depending upon the route by which thecomposition is to be administered, if applicable, and the effect whichthe practitioner desires the agent to have upon the nucleic acid orpolypeptide of the invention. Exemplary doses of a small moleculeinclude milligram or microgram amounts per kilogram of subject or sampleweight (e.g., about 1 microgram per kilogram to about 500 milligrams perkilogram, about 100 micrograms per kilogram to about 5 milligrams perkilogram, or about 1 microgram per kilogram to about 50 micrograms perkilogram). Exemplary doses of a protein or polypeptide include gram,milligram or microgram amounts per kilogram of subject or sample weight(e.g., about 1 microgram per kilogram to about 5 grams per kilogram,about 100 micrograms per kilogram to about 500 milligrams per kilogram,or about 1 milligram per kilogram to about 50 milligrams per kilogram).It is furthermore understood that appropriate doses of one of theseagents depend upon the potency of the agent with respect to theexpression or activity to be modulated. Such appropriate doses can bedetermined using the assays described herein. When one or more of theseagents is to be administered to an animal (e.g., a human) in order tomodulate expression or activity of a polypeptide or nucleic acid of theinvention, a physician, veterinarian, or researcher can, for example,prescribe a relatively low dose at first, subsequently increasing thedose until an appropriate response is obtained. In addition, it isunderstood that the specific dose level for any particular animalsubject will depend upon a variety of factors including the activity ofthe specific agent employed, the age, body weight, general health,gender, and diet of the subject, the time of administration, the routeof administration, the rate of excretion, any drug combination, and thedegree of expression or activity to be modulated.

[0437] A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), (topical), transmucosal, andrectal administration. Solutions or suspensions used for parenteral,intradermal, or subcutaneous application can include the followingcomponents: a sterile diluent such as water for injection, salinesolution, fixed oils, polyethylene glycols, glycerine, propylene glycolor other synthetic solvents; antibacterial agents such as benzyl alcoholor methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediamine-tetraacetic acid;buffers such as acetates, citrates or phosphates and agents for theadjustment of tonicity such as sodium chloride or dextrose. pH can beadjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampules,disposable syringes or multiple dose vials made of glass or plastic.

[0438] Pharmaceutical compositions suitable for injectable use includesterile aqueous solutions (where water soluble) or dispersions andsterile powders for the extemporaneous preparation of sterile injectablesolutions or dispersions. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorEL (BASF; Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the composition must be sterile and should be fluid to the extentthat easy syringability exists. It must be stable under the conditionsof manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyethylene glycol, and the like), and suitable mixturesthereof. The proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prevention of the action of microorganisms can be achieved by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride inthe composition. Prolonged absorption of the injectable compositions canbe brought about by including in the composition an agent which delaysabsorption, for example, aluminum monostearate and gelatin.

[0439] Sterile injectable solutions can be prepared by incorporating theactive compound (e.g., a polypeptide or antibody) in the required amountin an appropriate solvent with one or a combination of ingredientsenumerated above, as required, followed by filtered sterilization.Generally, dispersions are prepared by incorporating the active compoundinto a sterile vehicle which contains a basic dispersion medium, andthen incorporating the required other ingredients from those enumeratedabove. In the case of sterile powders for the preparation of sterileinjectable solutions, the preferred methods of preparation are vacuumdrying and freeze-drying which yields a powder of the active ingredientplus any additional desired ingredient from a previouslysterile-filtered solution thereof.

[0440] Oral compositions generally include an inert diluent or an ediblecarrier. They can be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound can be incorporated with excipients and used in the form oftablets, troches, or capsules. Oral compositions can also be preparedusing a fluid carrier for use as a mouthwash, wherein the compound inthe fluid carrier is applied orally and swished and expectorated orswallowed.

[0441] Pharmaceutically compatible binding agents, and/or adjuvantmaterials can be included as part of the composition. The tablets,pills, capsules, troches, and the like can contain any of the followingingredients, or compounds of a similar nature: a binder such asmicrocrystalline cellulose, gum tragacanth or gelatin; an excipient suchas starch or lactose, a disintegrating agent such as alginic acid,Primogel, or corn starch; a lubricant such as magnesium stearate orSterotes; a glidant such as colloidal silicon dioxide; a sweeteningagent such as sucrose or saccharin; or a flavoring agent such aspeppermint, methyl salicylate, or orange flavoring.

[0442] For administration by inhalation, the compounds are delivered inthe form of an aerosol spray from a pressurized container or dispenserwhich contains a suitable propellant, e.g., a gas such as carbondioxide, or a nebulizer.

[0443] Systemic administration can also be by transmucosal ortransdermal means. For transmucosal or transdermal administration,penetrants appropriate to the barrier to be permeated are used in theformulation. Such penetrants are generally known in the art, andinclude, for example, for transmucosal administration, detergents, bilesalts, and fusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the active compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

[0444] The compounds can also be prepared in the form of suppositories(e.g., with conventional suppository bases such as cocoa butter andother glycerides) or retention enemas for rectal delivery.

[0445] In one embodiment, the active compounds are prepared withcarriers that will protect the compound against rapid elimination fromthe body, such as a controlled release formulation, including implantsand microencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes which can be targeted to bind with virus-infectedcells using a monoclonal antibody which binds specifically with a viralantigen) can also be used as pharmaceutically acceptable carriers. Thesecan be prepared according to methods known to those skilled in the art,for example, as described in U.S. Pat. No. 4,522,811.

[0446] It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved, and the limitationsinherent in the art of compounding such an active compound for thetreatment of individuals.

[0447] For antibodies, the preferred dosage is 0.1 mg/kg to 100 mg/kg ofbody weight (generally 10 mg/kg to 20 mg/kg). If the antibody is to actin the brain, a dosage of 50 mg/kg to 100 mg/kg is usually appropriate.Generally, partially human antibodies and fully human antibodies have alonger half-life within the human body than other antibodies.Accordingly, lower dosages and less frequent administration is oftenpossible. Modifications such as lipidation can be used to stabilizeantibodies and to enhance uptake and tissue penetration (e.g., into thebrain). A method for lipidation of antibodies is described by Cruikshanket al. ((1997) J. Acquired Immune Deficiency Syndromes and HumanRetrovirology 14:193).

[0448] The nucleic acid molecules of the invention can be inserted intovectors and used as gene therapy vectors. Gene therapy vectors can bedelivered to a subject by, for example, intravenous injection, localadministration (U.S. Pat. No. 5,328,470), or by stereotactic injection(see, e.g., Chen et al. (1994) Proc. Natl. Acad. Sci. USA 91:3054-3057).The pharmaceutical preparation of the gene therapy vector can includethe gene therapy vector in an acceptable diluent, or can comprise a slowrelease matrix in which the gene delivery vehicle is imbedded.Alternatively, where the complete gene delivery vector can be producedintact from recombinant cells, e.g. retroviral vectors, thepharmaceutical preparation can include one or more cells which producethe gene delivery system.

[0449] The pharmaceutical compositions can be included in a container,pack, or dispenser together with instructions for administration.

[0450] Uses and Methods of the Invention

[0451] The nucleic acid molecules, proteins, protein homologs, andantibodies described herein can be used in one or more of the followingmethods: a) screening assays; b) detection assays (e.g., chromosomalmapping, tissue typing, forensic biology); c) predictive medicine (e.g.,diagnostic assays, prognostic assays, monitoring clinical trials, andpharmacogenomics); and d) methods of treatment (e.g., therapeutic andprophylactic). For example, polypeptides of the invention can to usedfor all of the purposes identified herein in portions of the disclosurerelating to individual types of protein of the invention (e.g., TANGO202 proteins, TANGO 234 proteins, TANGO 265 proteins, TANGO 273proteins, TANGO 286 proteins, TANGO 294 proteins, and INTERCEPT 296proteins). Polypeptides of the invention can also be used to modulatecellular proliferation, cellular differentiation, cellular adhesion, orsome combination of these. The isolated nucleic acid molecules of theinvention can be used to express proteins (e.g., via a recombinantexpression vector in a host cell in gene therapy applications), todetect mRNA (e.g., in a biological sample) or a genetic lesion, and tomodulate activity of a polypeptide of the invention. In addition, thepolypeptides of the invention can be used to screen drugs or compoundswhich modulate activity or expression of a polypeptide of the inventionas well as to treat disorders characterized by insufficient or excessiveproduction of a protein of the invention or production of a form of aprotein of the invention which has decreased or aberrant activitycompared to the wild type protein. In addition, the antibodies of theinvention can be used to detect and isolate a protein of the andmodulate activity of a protein of the invention.

[0452] This invention further pertains to novel agents identified by theabove-described screening assays and uses thereof for treatments asdescribed herein.

[0453] Screening Assays

[0454] The invention provides a method (also referred to herein as a“screening assay”) for identifying modulators, i.e., candidate or testcompounds or agents (e.g., peptides, peptidomimetics, small molecules orother drugs) which bind to polypeptide of the invention or have astimulatory or inhibitory effect on, for example, expression or activityof a polypeptide of the invention.

[0455] In one embodiment, the invention provides assays for screeningcandidate or test compounds which bind to or modulate the activity ofthe membrane-bound form of a polypeptide of the invention orbiologically active portion thereof. The test compounds of the presentinvention can be obtained using any of the numerous approaches incombinatorial library methods known in the art, including: biologicallibraries; spatially addressable parallel solid phase or solution phaselibraries; synthetic library methods requiring deconvolution; the“one-bead one-compound” library method; and synthetic library methodsusing affinity chromatography selection. The biological library approachis limited to peptide libraries, while the other four approaches areapplicable to peptide, non-peptide oligomer or small molecule librariesof compounds (Lam (1997) Anticancer Drug Des. 12:145).

[0456] Examples of methods for the synthesis of molecular libraries canbe found in the art, for example in: DeWitt et al. (1993) Proc. Natl.Acad. Sci. USA 90:6909; Erb et al. (1994) Proc. Natl. Acad. Sci. USA91:11422; Zuckermann et al. (1994). J. Med. Chem. 37:2678; Cho et al.(1993) Science 261:1303; Carrell et al. (1994) Angew. Chem. Int. Ed.Engl. 33:2059; Carell et al. (1994) Angew. Chem. Int. Ed. Engl. 33:2061;and Gallop et al. (1994) J. Med. Chem. 37:1233.

[0457] Libraries of compounds can be presented in solution (e.g.,Houghten (1992) Bio/Techniques 13:412-421), or on beads (Lam (1991)Nature 354:82-84), chips (Fodor (1993) Nature 364:555-556), bacteria(U.S. Pat. No. 5,223,409), spores (U.S. Pat. Nos. 5,571,698; 5,403,484;and 5,223,409), plasmids (Cull et al. (1992) Proc. Natl. Acad. Sci. USA89:1865-1869) or phage (Scott and Smith (1990) Science 249:386-390;Devlin (1990) Science 249:404-406; Cwirla et al. (1990) Proc. Natl.Acad. Sci. USA 87:6378-6382; and Felici (1991) J. Mol. Biol.222:301-310).

[0458] In one embodiment, an assay is a cell-based assay in which a cellwhich expresses a membrane-bound form of a polypeptide of the invention,or a biologically active portion thereof, on the cell surface iscontacted with a test compound and the ability of the test compound tobind to the polypeptide determined. The cell, for example, can be ayeast cell or a cell of mammalian origin. Determining the ability of thetest compound to bind to the polypeptide can be accomplished, forexample, by coupling the test compound with a radioisotope or enzymaticlabel such that binding of the test compound to the polypeptide orbiologically active portion thereof can be determined by detecting thelabeled compound in a complex. For example, test compounds can belabeled with ¹²⁵I, ³⁵S, ¹⁴C, or ³H, either directly or indirectly, andthe radioisotope detected by direct counting of radio-emission or byscintillation counting. Alternatively, test compounds can beenzymatically labeled with, for example, horseradish peroxidase,alkaline phosphatase, or luciferase, and the enzymatic label detected bydetermination of conversion of an appropriate substrate to product. In apreferred embodiment, the assay comprises contacting a cell whichexpresses a membrane-bound form of a polypeptide of the invention, or abiologically active portion thereof, on the cell surface with a knowncompound which binds the polypeptide to form an assay mixture,contacting the assay mixture with a test compound, and determining theability of the test compound to interact with the polypeptide, whereindetermining the ability of the test compound to interact with thepolypeptide comprises determining the ability of the test compound topreferentially bind to the polypeptide or a biologically active portionthereof as compared to the known compound.

[0459] In another embodiment, the assay involves assessment of anactivity characteristic of the polypeptide, wherein binding of the testcompound with the polypeptide or a biologically active portion thereofalters (i.e., increases or decreases) the activity of the polypeptide.

[0460] In another embodiment, an assay is a cell-based assay comprisingcontacting a cell expressing a membrane-bound form of a polypeptide ofthe invention, or a biologically active portion thereof, on the cellsurface with a test compound and determining the ability of the testcompound to modulate (e.g., stimulate or inhibit) the activity of thepolypeptide or biologically active portion thereof. Determining theability of the test compound to modulate the activity of the polypeptideor a biologically active portion thereof can be accomplished, forexample, by determining the ability of the polypeptide to bind to orinteract with a target molecule or to transport molecules across thecytoplasmic membrane.

[0461] Determining the ability of a polypeptide of the invention to bindto or interact with a target molecule can be accomplished by one of themethods described above for determining direct binding. As used herein,a “target molecule” is a molecule with which a selected polypeptide(e.g., a polypeptide of the invention binds or interacts with in nature,for example, a molecule on the surface of a cell which expresses theselected protein, a molecule on the surface of a second cell, a moleculein the extracellular milieu, a molecule associated with the internalsurface of a cell membrane or a cytoplasmic molecule. A target moleculecan be a polypeptide of the invention or some other polypeptide orprotein. For example, a target molecule can be a component of a signaltransduction pathway which facilitates transduction of an extracellularsignal (e.g., a signal generated by binding of a compound to apolypeptide of the invention) through the cell membrane and into thecell or a second intercellular protein which has catalytic activity or aprotein which facilitates the association of downstream signalingmolecules with a polypeptide of the invention. Determining the abilityof a polypeptide of the invention to bind to or interact with a targetmolecule can be accomplished by determining the activity of the targetmolecule. For example, the activity of the target molecule can bedetermined by detecting induction of a cellular second messenger of thetarget (e.g., an mRNA, intracellular Ca²⁺, diacylglycerol, IP3, and thelike), detecting catalytic/enzymatic activity of the target on anappropriate substrate, detecting the induction of a reporter gene (e.g.,a regulatory element that is responsive to a polypeptide of theinvention operably linked to a nucleic acid encoding a detectablemarker, e.g. luciferase), or detecting a cellular response, for example,cellular differentiation, or cell proliferation.

[0462] In yet another embodiment, an assay of the present invention is acell-free assay comprising contacting a polypeptide of the invention orbiologically active portion thereof with a test compound and determiningthe ability of the test compound to bind to the polypeptide orbiologically active portion thereof. Binding of the test compound to thepolypeptide can be determined either directly or indirectly as describedabove. In a preferred embodiment, the assay includes contacting thepolypeptide of the invention or biologically active portion thereof witha known compound which binds the polypeptide to form an assay mixture,contacting the assay mixture with a test compound, and determining theability of the test compound to interact with the polypeptide, whereindetermining the ability of the test compound to interact with thepolypeptide comprises determining the ability of the test compound topreferentially bind to the polypeptide or biologically active portionthereof as compared to the known compound.

[0463] In another embodiment, an assay is a cell-free assay comprisingcontacting a polypeptide of the invention or biologically active portionthereof with a test compound and determining the ability of the testcompound to modulate (e.g., stimulate or inhibit) the activity of thepolypeptide or biologically active portion thereof. Determining theability of the test compound to modulate the activity of the polypeptidecan be accomplished, for example, by determining the ability of thepolypeptide to bind to a target molecule by one of the methods describedabove for determining direct binding. In an alternative embodiment,determining the ability of the test compound to modulate the activity ofthe polypeptide can be accomplished by determining the ability of thepolypeptide of the invention to further modulate the target molecule Forexample, the catalytic activity, the enzymatic activity, or both, of thetarget molecule on an appropriate substrate can be determined aspreviously described.

[0464] In yet another embodiment, the cell-free assay comprisescontacting a polypeptide of the invention or biologically active portionthereof with a known compound which binds the polypeptide to form anassay mixture, contacting the assay mixture with a test compound, anddetermining the ability of the test compound to interact with thepolypeptide, wherein determining the ability of the test compound tointeract with the polypeptide comprises determining the ability of thepolypeptide to preferentially bind to or modulate the activity of atarget molecule.

[0465] The cell-free assays of the present invention are amenable to useof both a soluble form or the membrane-bound form of a polypeptide ofthe invention. In the case of cell-free assays comprising themembrane-bound form of the polypeptide, it can be desirable to utilize asolubilizing agent such that the membrane-bound form of the polypeptideis maintained in solution. Examples of such solubilizing agents includenon-ionic detergents such as n-octylglucoside, n-dodecylglucoside,n-octylmaltoside, octanoyl-N-methylglucamide,decanoyl-N-methylglucamide, Triton X-100, Triton X-114, Thesit,Isotridecypoly(ethylene glycol ether)n,3-[(3-cholamidopropyl)dimethylamminio]-1-propane sulfonate (CHAPS),3-[(3-cholamidopropyl)dimethylamminio]-2-hydroxy-1-propane sulfonate(CHAP S O), or N-dodecyl-N,N-dimethyl-3-ammonio-1-propane sulfonate.

[0466] In one or more embodiments of the above assay methods of thepresent invention, it can be desirable to immobilize either thepolypeptide of the invention or its target molecule to facilitateseparation of complexed from non-complexed forms of one or both of theproteins, as well as to accommodate automation of the assay. Binding ofa test compound to the polypeptide, or interaction of the polypeptidewith a target molecule in the presence and absence of a candidatecompound, can be accomplished in any vessel suitable for containing thereactants. Examples of such vessels include microtiter plates, testtubes, and micro-centrifuge tubes. In one embodiment, a fusion proteincan be provided which adds a domain that allows one or both of theproteins to be bound to a matrix. For example, glutathione-S-transferasefusion proteins or glutathione-S-transferase fusion proteins can beadsorbed onto glutathione SEPHAROSE™ beads (Sigma Chemical; St. Louis,Mo.) or glutathione derivatized microtiter plates, which are thencombined with the test compound or the test compound and either thenon-adsorbed target protein or A polypeptide of the invention, and themixture incubated under conditions conducive to complex formation (e.g.,at physiological conditions for salt and pH). Following incubation, thebeads or microtiter plate wells are washed to remove any unboundcomponents and complex formation is measured either directly orindirectly, for example, as described above. Alternatively, thecomplexes can be dissociated from the matrix, and the level of bindingor activity of the polypeptide of the invention can be determined usingstandard techniques.

[0467] Other techniques for immobilizing proteins on matrices can alsobe used in the screening assays of the invention. For example, eitherthe polypeptide of the invention or its target molecule can beimmobilized utilizing conjugation of biotin and streptavidin.Biotinylated polypeptide of the invention or target molecules can beprepared from biotin-NHS (N-hydroxy-succinimide) using techniques wellknown in the art (e.g., biotinylation kit, Pierce Chemicals; Rockford,Ill.), and immobilized in the wells of streptavidin-coated 96 wellplates (Pierce Chemical). Alternatively, antibodies reactive with thepolypeptide of the invention or target molecules but which do notinterfere with binding of the polypeptide of the invention to its targetmolecule can be derivatized to the wells of the plate, and unboundtarget or polypeptide of the invention trapped in the wells by antibodyconjugation. Methods for detecting such complexes, in addition to thosedescribed above for the GST-immobilized complexes, includeimmunodetection of complexes using antibodies reactive with thepolypeptide of the invention or target molecule, as well asenzyme-linked assays which rely on detecting an enzymatic activityassociated with the polypeptide of the invention or target molecule.

[0468] In another embodiment, modulators of expression of a polypeptideof the invention are identified in a method in which a cell is contactedwith a candidate compound and the expression of the selected mRNA orprotein (i.e., the mRNA or protein corresponding to a polypeptide ornucleic acid of the invention) in the cell is determined. The level ofexpression of the selected mRNA or protein in the presence of thecandidate compound is compared to the level of expression of theselected mRNA or protein in the absence of the candidate compound. Thecandidate compound can then be identified as a modulator of expressionof the polypeptide of the invention based on this comparison. Forexample, when expression of the selected mRNA or protein is greater(i.e., statistically significantly greater) in the presence of thecandidate compound than in its absence, the candidate compound isidentified as a stimulator of the selected mRNA or protein expression.Alternatively, when expression of the selected mRNA or protein is less(i.e., statistically significantly less) in the presence of thecandidate compound than in its absence, the candidate compound isidentified as an inhibitor of the selected mRNA or protein expression.The level of the selected mRNA or protein expression in the cells can bedetermined by methods described herein.

[0469] In yet another aspect of the invention, a polypeptide of theinventions can be used as “bait proteins” in a two-hybrid assay or threehybrid assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos et al. (1993)Cell 72:223-232; Madura et al. (1993) J. Biol. Chem. 268:12046-12054;Bartel et al. (1993) Bio/Techniques 14:920-924; Iwabuchi et al. (1993)Oncogene 8:1693-1696; and PCT Publication No. WO 94/10300), to identifyother proteins, which bind to or interact with the polypeptide of theinvention and modulate activity of the polypeptide of the invention.Such binding proteins are also likely to be involved in the propagationof signals by the polypeptide of the inventions as, for example,upstream or downstream elements of a signaling pathway involving thepolypeptide of the invention.

[0470] This invention further pertains to novel agents identified by theabove-described screening assays and uses thereof for treatments asdescribed herein.

[0471] Detection Assays

[0472] Portions or fragments of the cDNA sequences identified herein(and the corresponding complete gene sequences) can be used in numerousways as polynucleotide reagents. For example, these sequences can beused to: (i) map their respective genes on a chromosome and, thus,locate gene regions associated with genetic disease; (ii) identify anindividual from a minute biological sample (tissue typing); and (iii)aid in forensic identification of a biological sample. Theseapplications are described in the subsections below.

[0473] Chromosome Mapping

[0474] Once the sequence (or a portion of the sequence) of a gene hasbeen isolated, this sequence can be used to map the location of the geneon a chromosome. Accordingly, nucleic acid molecules described herein orfragments thereof, can be used to map the location of the correspondinggenes on a chromosome. The mapping of the sequences to chromosomes is animportant first step in correlating these sequences with genesassociated with disease.

[0475] Briefly, genes can be mapped to chromosomes by preparing PCRprimers (preferably 15-25 base pairs in length) from the sequence of agene of the invention. Computer analysis of the sequence of a gene ofthe invention can be used to rapidly select primers that do not spanmore than one exon in the genomic DNA, thus complicating theamplification process. These primers can then be used for PCR screeningof somatic cell hybrids containing individual human chromosomes. Onlythose hybrids containing the human gene corresponding to the genesequences will yield an amplified fragment. For a review of thistechnique, see D'Eustachio et al. ((1983) Science 220:919-924).

[0476] PCR mapping of somatic cell hybrids is a rapid procedure forassigning a particular sequence to a particular chromosome. Three ormore sequences can be assigned per day using a single thermal cycler.Using the nucleic acid sequences of the invention to designoligonucleotide primers, sub-localization can be achieved with panels offragments from specific chromosomes. Other mapping strategies which cansimilarly be used to map a gene to its chromosome include in situhybridization (described in Fan et al. (1990) Proc. Natl. Acad. Sci. USA87:6223-27), pre-screening with labeled flow-sorted chromosomes, andpre-selection by hybridization to chromosome specific cDNA libraries.Fluorescence in situ hybridization (FISH) of a DNA sequence to ametaphase chromosomal spread can further be used to provide a precisechromosomal location in one step. For a review of this technique, seeVerma et al. (Human Chromosomes: A Manual of Basic Techniques (PergamonPress, New York, 1988)).

[0477] Reagents for chromosome mapping can be used individually to marka single chromosome or a single site on that chromosome, or panels ofreagents can be used for marking multiple sites and/or multiplechromosomes. Reagents corresponding to non-coding regions of the genesactually are preferred for mapping purposes. Coding sequences are morelikely to be conserved within gene families, thus increasing the chanceof cross hybridizations during chromosomal mapping.

[0478] Once a sequence has been mapped to a precise chromosomallocation, the physical position of the sequence on the chromosome can becorrelated with genetic map data. (Such data are found, for example, inV. McKusick, Mendelian Inheritance in Man, available on-line throughJohns Hopkins University Welch Medical Library). The relationshipbetween genes and disease, mapped to the same chromosomal region, canthen be identified through linkage analysis (co-inheritance ofphysically adjacent genes), described in, e.g., Egeland et al. (1987)Nature 325:783-787.

[0479] Moreover, differences in the DNA sequences between individualsaffected and unaffected with a disease associated with a gene of theinvention can be determined. If a mutation is observed in some or all ofthe affected individuals but not in any unaffected individuals, then themutation is likely to be the causative agent of the particular disease.Comparison of affected and unaffected individuals generally involvesfirst looking for structural alterations in the chromosomes such asdeletions or translocations that are visible from chromosome spreads ordetectable using PCR based on that DNA sequence. Ultimately, completesequencing of genes from several individuals can be performed to confirmthe presence of a mutation and to distinguish mutations frompolymorphisms.

[0480] Furthermore, the nucleic acid sequences disclosed herein can beused to perform searches against “mapping databases”, e.g., BLAST-typesearch, such that the chromosome position of the gene is identified bysequence homology or identity with known sequence fragments which havebeen mapped to chromosomes.

[0481] In the instant case, the human gene for TANGO 265 is located onchromosome 1 between markers D1 S305 and D1S2635, and the human gene forTANGO 273 is located on chromosome 7 between markers D7S2467 andD7S2552.

[0482] In the instant case, the human gene for TANGO 286 exhibitssignificant amino acid homology with a region of the human chromosomeregion 22q12-13 genomic nucleotide sequence having GenBank Accessionnumber AL021937. Alignment of a 45 kilobase nucleotide sequence encodingTANGO 286 with AL021937, however, indicated the presence in TANGO 286 ofexons which differ from those disclosed in L021937 (pam120.mat scoringmatrix; gap penalties −12/−4). This region of chromosome 22 comprises animmunoglobulin lambda chain C (IGLC) pseudogene, the Ret fingerprotein-like 3 (RFPL3) and Ret finger protein-like 3 antisense (RFPL3S)genes, a gene encoding a novel immunoglobulin lambda chain V familyprotein, a novel gene encoding a protein similar both to mouse RGDSprotein (RALGDS, RALGEF, guanine nucleotide dissociation stimulator A)and to rabbit oncogene RSC, a novel gene encoding the human orthologueof worm F16A11.2 protein, a novel gene encoding a protein similar bothto BPI and to rabbit liposaccharide-binding protein, and a 5′-portion ofa novel gene. This region also comprises various ESTs, STSs, GSSs,genomic marker D22S1175, a ca repeat polymorphism and putative CpGislands.

[0483] A polypeptide and fragments and sequences thereof and antibodieswhich bind specifically with such polypeptides/fragments can be used tomap the location of the gene encoding the polypeptide on a chromosome.This mapping can be performed by specifically detecting the presence ofthe polypeptide/fragments in members of a panel of somatic cell hybridsbetween cells obtained from a first species of animal from which theprotein originates and cells obtained from a second species of animal,determining which somatic cell hybrid(s) expresses the polypeptide, andnoting the chromosome(s) of the first species of animal that itcontains. For examples of this technique (see Pajunen et al., 1988,Cytogenet. Cell Genet. 47:37-41 and Van Keuren et al., 1986, Hum. Genet.74:34-40). Alternatively, the presence of the polypeptide in the somaticcell hybrids can be determined by assaying an activity or property ofthe polypeptide (e.g., enzymatic activity, as described inBordelon-Riser et al., 1979, Som. Cell Genet. 5:597-613 and Owerbach etal., 1978, Proc. Natl. Acad. Sci. USA 75:5640-5644).

[0484] In the instant case, the human gene for TANGO 234 proteinindicated that the gene is located at chromosomal location h12p13.Flanking chromosomal markers include WI-6980 and GATA8A09.43. Nearbyhuman loci include IBD2 (inflammatory bowel disease 2), FPF (familialperiodic fever), and HPDR2 (hypophosphatemia vitamin D resistant rickets2). Nearby genes are KLRC (killer cell receptor cluster), DRPLA(dentatorubro-pallidoluysian atrophy), GAPD (glyceraldehyde-3-phosphate)dehydrogenase, and PXR1 peroxisome receptor 1). This region is syntenicto mouse chromosome mo6. Murine chromosomal mapping indicated that themurine orthologue is located near the scr (scruffy) locus. Nearby mousegenes include drpla (dentatorubral phillidoluysian atrophy), prp(proline rich protein), and kap (kidney androgen regulated protein).

[0485] Tissue Typing

[0486] The nucleic acid sequences of the present invention can also beused to identify individuals from minute biological samples. The UnitedStates military, for example, is considering the use of restrictionfragment length polymorphism (RFLP) for identification of its personnel.In this technique, an individual's genomic DNA is digested with one ormore restriction enzymes, and probed on a Southern blot to yield uniquebands for identification. This method does not suffer from the currentlimitations of “Dog Tags” which can be lost, switched, or stolen, makingpositive identification difficult. The sequences of the presentinvention are useful as additional DNA markers for RFLP (described inU.S. Pat. No. 5,272,057).

[0487] Furthermore, the sequences of the present invention can be usedto provide an alternative technique which determines the actualbase-by-base DNA sequence of selected portions of an individual'sgenome. Thus, the nucleic acid sequences described herein can be used toprepare two PCR primers from the 5′ and 3′ ends of the sequences. Theseprimers can then be used to amplify an individual's DNA and subsequentlysequence it.

[0488] Panels of corresponding DNA sequences from individuals, preparedin this manner, can provide unique individual identifications, as eachindividual will have a unique set of such DNA sequences due to allelicdifferences. The sequences of the present invention can be used toobtain such identification sequences from individuals and from tissue.The nucleic acid sequences of the invention uniquely represent portionsof the human genome. Allelic variation occurs to some degree in thecoding regions of these sequences, and to a greater degree in thenon-coding regions. It is estimated that allelic variation betweenindividual humans occurs with a frequency of about once per each 500bases. Each of the sequences described herein can, to some degree, beused as a standard against which DNA from an individual can be comparedfor identification purposes. Because greater numbers of polymorphismsoccur in the non-coding regions, fewer sequences are necessary todifferentiate individuals. The non-coding sequences of any of SEQ IDNOs: 1, 9, 17, 25, 33, 45, and 53 can comfortably provide positiveindividual identification with a panel of perhaps 10 to 1,000 primerswhich each yield a non-coding amplified sequence of 100 bases. Ifpredicted coding sequences, such as those in any of SEQ ID NOs: 2, 10,18, 26, 34, 46, and 54 are used, a more appropriate number of primersfor positive individual identification would be 500-2,000.

[0489] If a panel of reagents from the nucleic acid sequences describedherein is used to generate a unique identification database for anindividual, those same reagents can later be used to identify tissuefrom that individual. Using the unique identification database, positiveidentification of the individual, living or dead, can be made fromextremely small tissue samples.

[0490] Use of Partial Gene Sequences in Forensic Biology

[0491] DNA-based identification techniques can also be used in forensicbiology. Forensic biology is a scientific field employing genetic typingof biological evidence found at a crime scene as a means for positivelyidentifying, for example, a perpetrator of a crime. To make such anidentification, PCR technology can be used to amplify DNA sequencestaken from very small biological samples such as tissues, e.g., hair orskin, or body fluids, e.g., blood, saliva, or semen found at a crimescene. The amplified sequence can then be compared to a standard,thereby allowing identification of the origin of the biological sample.

[0492] The sequences of the present invention can be used to providepolynucleotide reagents, e.g., PCR primers, targeted to specific loci inthe human genome, which can enhance the reliability of DNA-basedforensic identifications by, for example, providing another“identification marker” (i.e., another DNA sequence that is unique to aparticular individual). As mentioned above, actual base sequenceinformation can be used for identification as an accurate alternative topatterns formed by restriction enzyme generated fragments. Sequencestargeted to non-coding regions are particularly appropriate for this useas greater numbers of polymorphisms occur in the non-coding regions,making it easier to differentiate individuals using this technique.Examples of polynucleotide reagents include the nucleic acid sequencesof the invention or portions thereof, e.g., fragments derived fromnon-coding regions having a length of at least 20 or 30 bases.

[0493] The nucleic acid sequences described herein can further be usedto provide polynucleotide reagents, e.g., labeled or labelable probeswhich can be used in, for example, an in situ hybridization technique,to identify a specific tissue, e.g., brain tissue. This can be veryuseful in cases where a forensic pathologist is presented with a tissueof unknown origin. Panels of such probes can be used to identify tissueby species and/or by organ type.

[0494] Predictive Medicine

[0495] The present invention also pertains to the field of predictivemedicine in which diagnostic assays, prognostic assays,pharmacogenomics, and monitoring clinical trials are used for prognostic(predictive) purposes to thereby treat an individual prophylactically.Accordingly, one aspect of the present invention relates to diagnosticassays for determining expression of a polypeptide or nucleic acid ofthe invention and/or activity of a polypeptide of the invention (e.g.,expression or activity of one of TANGO 202, TANGO 234, TANGO 265, TANGO273, TANGO 286, TANGO 294, or INTERCEPT 296 genes or proteins), in thecontext of a biological sample (e.g., blood, serum, cells, tissue) tothereby determine whether an individual is afflicted with a disease ordisorder, or is at risk of developing a disorder, associated withaberrant expression or activity of a polypeptide of the invention. Theinvention also provides for prognostic (or predictive) assays fordetermining whether an individual is at risk of developing a disorderassociated with aberrant expression or activity of a polypeptide of theinvention. For example, mutations in a gene of the invention can beassayed in a biological sample. Such assays can be used for prognosticor predictive purpose to thereby prophylactically treat an individualprior to the onset of a disorder characterized by or associated withaberrant expression or activity of a polypeptide of the invention.

[0496] As an alternative to making determinations based on the absoluteexpression level of a selected gene, determinations can be based onnormalized expression levels of the gene. A gene expression level isnormalized by correcting the absolute expression level of the gene(e.g., a TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO294, or INTERCEPT 296 gene as described herein) by comparing itsexpression to expression of a gene for which expression is not believedto be co-regulated with the gene of interest, e.g., a housekeeping genethat is constitutively expressed. Suitable genes for normalizationinclude housekeeping genes such as the actin gene. Such normalizationallows comparison of the expression level in one sample, e.g., a patientsample, with the expression level in another sample, e.g., a sampleobtained from a patient known not to be afflicted with a disease orcondition, or between samples obtained from different sources.

[0497] Alternatively, the expression level can be assessed as a relativeexpression level. To assess a relative expression level for a gene(e.g., a TANGO 202, TANGO 234, TANGO 265, TANGO 273, TANGO 286, TANGO294, or INTERCEPT 296 gene, as described herein), the level ofexpression of the gene is determined for 10 or more samples (preferably50 or more samples) of different isolates of cells in which the gene isbelieved to be expressed, prior to assessing the level of expression ofthe gene in the sample of interest. The mean expression level of thegene detected in the large number of samples is determined, and thisvalue is used as a baseline expression level for the gene. Theexpression level of the gene assessed in the test sample (i.e., itsabsolute level of expression) is divided by the mean expression value toyield a relative expression level. Such a method can identify tissues orindividuals which are afflicted with a disorder associated with aberrantexpression of a gene of the invention.

[0498] Preferably, the samples used in the baseline determination aregenerated either using cells obtained from a tissue or individual knownto be afflicted with a disorder (e.g., a disorder associated withaberrant expression of one of the TANGO 202, TANGO 234, TANGO 265, TANGO273, TANGO 286, TANGO 294, or INTERCEPT 296 genes) or using cellsobtained from a tissue or individual known not to be afflicted with thedisorder. Alternatively, levels of expression of these genes in tissuesor individuals known to be or not to be afflicted with the disorder canbe used to assess whether the aberrant expression of the gene isassociated with the disorder (e.g., with onset of the disorder, or as asymptom of the disorder over time).

[0499] Another aspect of the invention pertains to monitoring theinfluence of agents (e.g., drugs or other compounds) on the expressionor activity of one or more of TANGO 202, TANGO 234, TANGO 265, TANGO273, TANGO 286, TANGO 294, and INTERCEPT 296 in clinical trials. Theseand other agents are described in further detail in the followingsections.

[0500] Diagnostic Assays

[0501] An exemplary method for detecting the presence or absence of apolypeptide or nucleic acid of the invention in a biological sampleinvolves obtaining a biological sample from a test subject andcontacting the biological sample with a compound or an agent capable ofdetecting a polypeptide or nucleic acid (e.g., mRNA, genomic DNA) of theinvention such that the presence of a polypeptide or nucleic acid of theinvention is detected in the biological sample. A preferred agent fordetecting mRNA or genomic DNA encoding a polypeptide of the invention isa labeled nucleic acid probe capable of hybridizing to mRNA or genomicDNA encoding a polypeptide of the invention. The nucleic acid probe canbe, for example, a full-length cDNA, such as the nucleic acid of any ofSEQ ID NOs: 1, 9, 17, 25, 33, 45, 53, 67, and 72, or a portion thereof,such as an oligonucleotide of at least 15, 30, 50, 100, 250 or 500nucleotides in length and sufficient to specifically hybridize understringent conditions to a mRNA or genomic DNA encoding a polypeptide ofthe invention. Other suitable probes for use in the diagnostic assays ofthe invention are described herein.

[0502] A preferred agent for detecting a polypeptide of the invention isan antibody capable of binding to a polypeptide of the invention,preferably an antibody with a detectable label Antibodies can bepolyclonal, or more preferably, monoclonal. An intact antibody, or afragment thereof (e.g., Fab or F(ab′)₂) can be used. The term “labeled”,with regard to the probe or antibody, is intended to encompass directlabeling of the probe or antibody by coupling (i.e., physically linking)a detectable substance to the probe or antibody, as well as indirectlabeling of the probe or antibody by reactivity with another reagentthat is directly labeled. Examples of indirect labeling includedetection of a primary antibody using a fluorescently labeled secondaryantibody and end-labeling of a DNA probe with biotin such that it can bedetected with fluorescently labeled streptavidin. The term “biologicalsample” is intended to include tissues, cells and biological fluidsisolated from a subject, as well as tissues, cells and fluids presentwithin a subject. That is, the detection method of the invention can beused to detect mRNA, protein, or genomic DNA in a biological sample invitro as well as in vivo. For example, in vitro techniques for detectionof mRNA include Northern hybridizations and in situ hybridizations. Invitro techniques for detection of a polypeptide of the invention includeenzyme linked immunosorbent assays (ELISAs), Western blots,immunoprecipitations and immunofluorescence. In vitro techniques fordetection of genomic DNA include Southern hybridizations. Furthermore,in vivo techniques for detection of a polypeptide of the inventioninclude introducing into a subject a labeled antibody directed againstthe polypeptide. For example, the antibody can be labeled with aradioactive marker whose presence and location in a subject can bedetected by standard imaging techniques.

[0503] In one embodiment, the biological sample contains proteinmolecules from the test subject. Alternatively, the biological samplecan contain mRNA molecules from the test subject or genomic DNAmolecules from the test subject. A preferred biological sample is aperipheral blood leukocyte sample isolated by conventional means from asubject.

[0504] In another embodiment, the methods further involve obtaining acontrol biological sample from a control subject, contacting the controlsample with a compound or agent capable of detecting a polypeptide ofthe invention or mRNA or genomic DNA encoding a polypeptide of theinvention, such that the presence of the polypeptide or mRNA or genomicDNA encoding the polypeptide is detected in the biological sample, andcomparing the presence of the polypeptide or mRNA or genomic DNAencoding the polypeptide in the control sample with the presence of thepolypeptide or mRNA or genomic DNA encoding the polypeptide in the testsample.

[0505] The invention also encompasses kits for detecting the presence ofa polypeptide or nucleic acid of the invention in a biological sample (atest sample). Such kits can be used to determine if a subject issuffering from or is at increased risk of developing a disorderassociated with aberrant expression of a polypeptide of the invention(e.g., one of the disorders described in the section of this disclosurewherein the individual polypeptide of the invention is discussed). Forexample, the kit can comprise a labeled compound or agent capable ofdetecting the polypeptide or mRNA encoding the polypeptide in abiological sample and means for determining the amount of thepolypeptide or mRNA in the sample (e.g., an antibody which binds thepolypeptide or an oligonucleotide probe which binds to DNA or mRNAencoding the polypeptide). Kits can also include instructions forobserving that the tested subject is suffering from or is at risk ofdeveloping a disorder associated with aberrant expression of thepolypeptide if the amount of the polypeptide or mRNA encoding thepolypeptide is above or below a normal level.

[0506] For antibody-based kits, the kit can comprise, for example: (1) afirst antibody (e.g., attached to a solid support) which binds to apolypeptide of the invention; and, optionally, (2) a second, differentantibody which binds to either the polypeptide or the first antibody andis conjugated to a detectable agent.

[0507] For oligonucleotide-based kits, the kit can comprise, forexample: (1) an oligonucleotide, e.g., a detectably labeledoligonucleotide, which hybridizes to a nucleic acid sequence encoding apolypeptide of the invention or (2) a pair of primers useful foramplifying a nucleic acid molecule encoding a polypeptide of theinvention. The kit can also comprise, e.g., a buffering agent, apreservative, or a protein stabilizing agent. The kit can also comprisecomponents necessary for detecting the detectable agent (e.g., an enzymeor a substrate). The kit can also contain a control sample or a seriesof control samples which can be assayed and compared to the test samplecontained. Each component of the kit is usually enclosed within anindividual container and all of the various containers are within asingle package along with instructions for observing whether the testedsubject is suffering from or is at risk of developing a disorderassociated with aberrant expression of the polypeptide.

[0508] Prognostic Assays

[0509] The methods described herein can furthermore be utilized asdiagnostic or prognostic assays to identify subjects having or at riskof developing a disease or disorder associated with aberrant expressionor activity of a polypeptide of the invention. For example, the assaysdescribed herein, such as the preceding diagnostic assays or thefollowing assays, can be utilized to identify a subject having or atrisk of developing a disorder associated with aberrant expression oractivity of a polypeptide of the invention (e.g., one of the disordersdescribed in the section of this disclosure wherein the individualpolypeptide of the invention is discussed). Alternatively, theprognostic assays can be utilized to identify a subject having or atrisk for developing such a disease or disorder. Thus, the presentinvention provides a method in which a test sample is obtained from asubject and a polypeptide or nucleic acid (e.g., mRNA, genomic DNA) ofthe invention is detected, wherein the presence of the polypeptide ornucleic acid is diagnostic for a subject having or at risk of developinga disease or disorder associated with aberrant expression or activity ofthe polypeptide. As used herein, a “test sample” refers to a biologicalsample obtained from a subject of interest. For example, a test samplecan be a biological fluid (e.g., serum), cell sample, or tissue.

[0510] Furthermore, the prognostic assays described herein can be usedto determine whether a subject can be administered an agent (e.g., anagonist, antagonist, peptidomimetic, protein, peptide, nucleic acid,small molecule, or other drug candidate) to treat a disease or disorderassociated with aberrant expression or activity of a polypeptide of theinvention. For example, such methods can be used to determine whether asubject can be effectively treated with a specific agent or class ofagents (e.g., agents of a type which decrease activity of thepolypeptide). Thus, the present invention provides methods fordetermining whether a subject can be effectively treated with an agentfor a disorder associated with aberrant expression or activity of apolypeptide of the invention in which a test sample is obtained and thepolypeptide or nucleic acid encoding the polypeptide is detected (e.g.,wherein the presence of the polypeptide or nucleic acid is diagnosticfor a subject that can be administered the agent to treat a disorderassociated with aberrant expression or activity of the polypeptide).

[0511] The methods of the invention can also be used to detect geneticlesions or mutations in a gene of the invention, thereby determining ifa subject with the lesioned gene is at risk for a disorder characterizedaberrant expression or activity of a polypeptide of the invention. Inpreferred embodiments, the methods include detecting, in a sample ofcells from the subject, the presence or absence of a genetic lesion ormutation characterized by at least one of an alteration affecting theintegrity of a gene encoding the polypeptide of the invention, or themis-expression of the gene encoding the polypeptide of the invention.For example, such genetic lesions or mutations can be detected byascertaining the existence of at least one of: 1) a deletion of one ormore nucleotides from the gene; 2) an addition of one or morenucleotides to the gene; 3) a substitution of one or more nucleotides ofthe gene; 4) a chromosomal rearrangement of the gene; 5) an alterationin the level of a messenger RNA transcript of the gene; 6) an aberrantmodification of the gene, such as of the methylation pattern of thegenomic DNA; 7) the presence of a non-wild type splicing pattern of amessenger RNA transcript of the gene; 8) a non-wild type level of theprotein encoded by the gene; 9) an allelic loss of the gene; and 10) aninappropriate post-translational modification of the protein encoded bythe gene. As described herein, there are a large number of assaytechniques known in the art which can be used for detecting lesions in agene.

[0512] In certain embodiments, detection of the lesion involves the useof a probe/primer in a polymerase chain reaction (PCR) (see, e.g., U.S.Pat. Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or,alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegranet al. (1988) Science 241:1077-1080; and Nakazawa et al. (1994) Proc.Natl. Acad. Sci. USA 91:360-364), the latter of which can beparticularly useful for detecting point mutations in a gene (see, e.g.,Abravaya et al. (1995) Nucleic Acids Res. 23:675-682). This method caninclude the steps of collecting a sample of cells from a patient,isolating nucleic acid (e.g., genomic, mRNA or both) from the cells ofthe sample, contacting the nucleic acid sample with one or more primerswhich specifically hybridize to the selected gene under conditions suchthat hybridization and amplification of the gene (if present) occurs,and detecting the presence or absence of an amplification product, ordetecting the size of the amplification product and comparing the lengthto a control sample. PCR and/or LCR can be desirable to use as apreliminary amplification step in conjunction with any of the techniquesused for detecting mutations described herein.

[0513] Alternative amplification methods include: self-sustainedsequence replication (Guatelli et al. (1990) Proc. Natl. Acad. Sci. USA87:1874-1878), transcriptional amplification system (Kwoh, et al. (1989)Proc. Natl. Acad. Sci. USA 86:1173-1177), Q-Beta Replicase (Lizardi etal. (1988) Bio/Technology 6:1197), or any other nucleic acidamplification method, followed by the detection of the amplifiedmolecules using techniques well known to those of skill in the art.These detection schemes are especially useful for the detection ofnucleic acid molecules if such molecules are present in very lownumbers.

[0514] In an alternative embodiment, mutations in a selected gene from asample cell can be identified by alterations in restriction enzymecleavage patterns. For example, sample and control DNA is isolated,(optionally) amplified, digested with one or more restrictionendonucleases, and fragment length sizes are determined by gelelectrophoresis and compared. Differences in fragment length sizesbetween sample and control DNA indicates mutations in the sample DNA.Moreover, the use of sequence specific ribozymes (see, e.g., U.S. Pat.No. 5,498,531) can be used to score for the presence of specificmutations by development or loss of a ribozyme cleavage site.

[0515] In other embodiments, genetic mutations can be identified byhybridizing a sample and control nucleic acids, e.g., DNA or RNA, tohigh density arrays containing hundreds or thousands of oligonucleotidesprobes (Cronin et al. (1996) Human Mutation 7:244-255; Kozal et al.(1996) Nature Medicine 2:753-759). For example, genetic mutations can beidentified in two-dimensional arrays containing light-generated DNAprobes as described in Cronin et al., supra. Briefly, a firsthybridization array of probes can be used to scan through long stretchesof DNA in a sample and control to identify base changes between thesequences by making linear arrays of sequential overlapping probes. Thisstep allows the identification of point mutations. This step is followedby a second hybridization array that allows the characterization ofspecific mutations by using smaller, specialized probe arrayscomplementary to all variants or mutations detected. Each mutation arrayis composed of parallel probe sets, one complementary to the wild-typegene and the other complementary to the mutant gene.

[0516] In yet another embodiment, any of a variety of sequencingreactions known in the art can be used to directly sequence the selectedgene and detect mutations by comparing the sequence of the samplenucleic acids with the corresponding wild-type (control) sequence.Examples of sequencing reactions include those based on techniquesdeveloped by Maxim and Gilbert ((1977) Proc. Natl. Acad. Sci. USA74:560) or Sanger ((1977) Proc. Natl. Acad. Sci. USA 74:5463). It isalso contemplated that any of a variety of automated sequencingprocedures can be utilized when performing the diagnostic assays ((1995)Bio/Techniques 19:448), including sequencing by mass spectrometry (see,e.g., PCT Publication No. WO 94/16101; Cohen et al. (1996) Adv.Chromatogr. 36:127-162; and Griffin et al. (1993) Appl. Biochem.Biotechnol. 38:147-159).

[0517] Other methods for detecting mutations in a selected gene includemethods in which protection from cleavage agents is used to detectmismatched bases in RNA/RNA or RNA/DNA heteroduplexes (Myers et al.(1985) Science 230:1242). In general, the technique of mismatch cleavageentails providing heteroduplexes formed by hybridizing (labeled) RNA orDNA containing the wild-type sequence with potentially mutant RNA or DNAobtained from a tissue sample. The double-stranded duplexes are treatedwith an agent which cleaves single-stranded regions of the duplex suchas which will exist due to base pair mismatches between the control andsample strands. RNA/DNA duplexes can be treated with RNase to digestmismatched regions, and DNA/DNA hybrids can be treated with S1 nucleaseto digest mismatched regions.

[0518] In other embodiments, either DNA/DNA or RNA/DNA duplexes can betreated with hydroxylamine or osmium tetroxide and with piperidine inorder to digest mismatched regions. After digestion of the mismatchedregions, the resulting material is then separated by size on denaturingpolyacrylamide gels to determine the site of mutation. See, e.g., Cottonet al. (1988) Proc. Natl. Acad. Sci. USA 85:4397; Saleeba et al. (1992)Methods Enzymol. 217:286-295. In a preferred embodiment, the control DNAor RNA can be labeled for detection.

[0519] In still another embodiment, the mismatch cleavage reactionemploys one or more proteins that recognize mismatched base pairs indouble-stranded DNA (so called DNA mismatch repair enzymes) in definedsystems for detecting and mapping point mutations in cDNAs obtained fromsamples of cells. For example, the mutY enzyme of E. coli cleaves A atG/A mismatches and the thymidine DNA glycosylase from HeLa cells cleavesT at G/T mismatches (Hsu et al. (1994) Carcinogenesis 15:1657-1662).According to an exemplary embodiment, a probe based on a selectedsequence, e.g., a wild-type sequence, is hybridized to a cDNA or otherDNA product from a test cell(s). The duplex is treated with a DNAmismatch repair enzyme, and the cleavage products, if any, can bedetected from electrophoresis protocols or the like. See, e.g., U.S.Pat. No. 5,459,039.

[0520] In other embodiments, alterations in electrophoretic mobilitywill be used to identify mutations in genes. For example, single strandconformation polymorphism (SSCP) can be used to detect differences inelectrophoretic mobility between mutant and wild type nucleic acids(Orita et al. (1989) Proc. Natl. Acad. Sci. USA 86:2766; see also Cotton(1993) Mutat. Res. 285:125-144; Hayashi (1992) Genet. Anal. Tech. Appl.9:73-79). Single-stranded DNA fragments of sample and control nucleicacids will be denatured and allowed to re-nature. The secondarystructure of single-stranded nucleic acids varies according to sequence,and the resulting alteration in electrophoretic mobility enables thedetection of even a single base change. The DNA fragments can be labeledor detected with labeled probes. The sensitivity of the assay can beenhanced by using RNA (rather than DNA), in which the secondarystructure is more sensitive to a change in sequence. In a preferredembodiment, the subject method utilizes heteroduplex analysis toseparate double stranded heteroduplex molecules on the basis of changesin electrophoretic mobility (Keen et al. (1991) Trends Genet. 7:5).

[0521] In yet another embodiment, the movement of mutant or wild-typefragments in polyacrylamide gels containing a gradient of denaturant isassayed using denaturing gradient gel electrophoresis (DGGE) (Myers etal. (1985) Nature 313:495). When DGGE is used as the method of analysis,DNA will be modified to insure that it does not completely denature, forexample by adding a ‘GC clamp’ of approximately 40 base pairs ofhigh-melting GC-rich DNA by PCR. In a further embodiment, a temperaturegradient is used in place of a denaturing gradient to identifydifferences in the mobility of control and sample DNA (Rosenbaum andReissner (1987) Biophys. Chem. 265:12753).

[0522] Examples of other techniques for detecting point mutationsinclude, but are not limited to, selective oligonucleotidehybridization, selective amplification, or selective primer extension.For example, oligonucleotide primers can be prepared in which the knownmutation is placed centrally and then hybridized to target DNA underconditions which permit hybridization only if a perfect match is found(Saiki et al. (1986) Nature 324:163); Saiki et al. (1989) Proc. Natl.Acad. Sci. USA 86:6230). Such allele specific oligonucleotides arehybridized to PCR amplified target DNA or a number of differentmutations when the oligonucleotides are attached to the hybridizingmembrane and hybridized with labeled target DNA.

[0523] Alternatively, allele specific amplification technology whichdepends on selective PCR amplification can be used in conjunction withthe instant invention. Oligonucleotides used as primers for specificamplification can carry the mutation of interest in the center of themolecule (so that amplification depends on differential hybridization;Gibbs et al. (1989) Nucleic Acids Res. 17:2437-2448) or at the extreme3′ end of one primer where, under appropriate conditions, mismatchingcan prevent or reduce polymerase extension (Prossner (1993) Tibtech11:238). In addition, it can be desirable to introduce a novelrestriction site in the region of the mutation to create cleavage-baseddetection (Gasparini et al. (1992) Mol. Cell Probes 6:1). Amplificationcan also be performed using Taq ligase for amplification (Barany (1991)Proc. Natl. Acad. Sci. USA 88:189). In such cases, ligation will occuronly if there is a perfect match at the 3′ end of the 5′ sequence makingit possible to detect the presence of a known mutation at a specificsite by looking for the presence or absence of amplification.

[0524] The methods described herein can be performed, for example, usingpre-packaged diagnostic kits comprising at least one probe nucleic acidor antibody reagent described herein, which can be conveniently used,e.g., in clinical settings to diagnose patients exhibiting symptoms orfamily history of a disease or illness involving a gene encoding apolypeptide of the invention. Furthermore, any cell type or tissue,preferably peripheral blood leukocytes, in which the polypeptide of theinvention is expressed can be utilized in the prognostic assaysdescribed herein.

[0525] Pharmacogenomics

[0526] Agents, or modulators which have a stimulatory or inhibitoryeffect on activity or expression of a polypeptide of the invention asidentified by a screening assay described herein can be administered toindividuals to treat (prophylactically or therapeutically) disordersassociated with aberrant activity of the polypeptide. In conjunctionwith such treatment, the pharmacogenomics (i.e., the study of therelationship between an individual's genotype and that individual'sresponse to a foreign compound or drug) of the individual may beconsidered. Differences in metabolism of therapeutics can lead to severetoxicity or therapeutic failure by altering the relation between doseand blood concentration of the pharmacologically active drug. Thus, thepharmacogenomics of the individual permits the selection of effectiveagents (e.g., drugs) for prophylactic or therapeutic treatments based ona consideration of the individual's genotype. Such pharmacogenomics canfurther be used to determine appropriate dosages and therapeuticregimens. Accordingly, the activity of a polypeptide of the invention,expression of a nucleic acid of the invention, or mutation content of agene of the invention in an individual can be determined to therebyselect appropriate agent(s) for therapeutic or prophylactic treatment ofthe individual.

[0527] Pharmacogenomics deals with clinically significant hereditaryvariations in the response to drugs due to altered drug disposition andabnormal action in affected persons. See, e.g., Linder (1997) Clin.Chem. 43(2):254-266. In general, two types of pharmacogenetic conditionscan be differentiated. Genetic conditions transmitted as a single factoraltering the way drugs act on the body are referred to as “altered drugaction.” Genetic conditions transmitted as single factors altering theway the body acts on drugs are referred to as “altered drug metabolism”.These pharmacogenetic conditions can occur either as rare defects or aspolymorphisms. For example, glucose-6-phosphate dehydrogenase (G6PD)deficiency is a common inherited enzymopathy in which the main clinicalcomplication is hemolysis after ingestion of oxidant drugs(anti-malarials, sulfonamides, analgesics, nitrofurans) and consumptionof fava beans.

[0528] As an illustrative embodiment, the activity of drug metabolizingenzymes is a major determinant of both the intensity and duration ofdrug action. The discovery of genetic polymorphisms of drug metabolizingenzymes (e.g., N-acetyltransferase 2 (NAT 2) and cytochrome P450 enzymesCYP2D6 and CYP2C19) has provided an explanation as to why some patientsdo not obtain the expected drug effects or show exaggerated drugresponse and serious toxicity after taking the standard and safe dose ofa drug. These polymorphisms are expressed in two phenotypes in thepopulation, the extensive metabolizer (EM) and poor metabolizer (PM).The prevalence of PM is different among different populations. Forexample, the gene coding for CYP2D6 is highly polymorphic and severalmutations have been identified in PM, which all lead to the absence offunctional CYP2D6. Poor metabolizers of CYP2D6 and CYP2C19 quitefrequently experience exaggerated drug response and side effects whenthey receive standard doses. If a metabolite is the active therapeuticmoiety, a PM will show no therapeutic response, as demonstrated for theanalgesic effect of codeine mediated by its CYP2D6-formed metabolitemorphine. The other extreme are the so called ultra-rapid metabolizerswho do not respond to standard doses. Recently, the molecular basis ofultra-rapid metabolism has been identified to be due to CYP2D6 geneamplification.

[0529] Thus, the activity of a polypeptide of the invention, expressionof a nucleic acid encoding the polypeptide, or mutation content of agene encoding the polypeptide in an individual can be determined tothereby select appropriate agent(s) for therapeutic or prophylactictreatment of the individual. In addition, pharmacogenetic studies can beused to apply genotyping of polymorphic alleles encodingdrug-metabolizing enzymes to the identification of an individual's drugresponsiveness phenotype. This knowledge, when applied to dosing or drugselection, can avoid adverse reactions or therapeutic failure and thusenhance therapeutic or prophylactic efficiency when treating a subjectwith a modulator of activity or expression of the polypeptide, such as amodulator identified by one of the exemplary screening assays describedherein.

[0530] Monitoring of Effects During Clinical Trials

[0531] Monitoring the influence of agents (e.g., drug compounds) on theexpression or activity of a polypeptide of the invention (e.g., theability to modulate aberrant cell proliferation chemotaxis, and/ordifferentiation) can be applied not only in basic drug screening, butalso in clinical trials. For example, the effectiveness of an agent, asdetermined by a screening assay as described herein, to increase geneexpression, protein levels, or protein activity, can be monitored inclinical trials of subjects exhibiting decreased gene expression,protein levels, or protein activity. Alternatively, the effectiveness ofan agent, as determined by a screening assay, to decrease geneexpression, protein levels or protein activity, can be monitored inclinical trials of subjects exhibiting increased gene expression,protein levels, or protein activity. In such clinical trials, expressionor activity of a polypeptide of the invention and preferably, that ofother polypeptide that have been implicated in for example, a cellularproliferation disorder, can be used as a marker of the immuneresponsiveness of a particular cell.

[0532] For example, and not by way of limitation, genes, including thoseof the invention, that are modulated in cells by treatment with an agent(e.g., compound, drug or small molecule) which modulates activity orexpression of a polypeptide of the invention (e.g., as identified in ascreening assay described herein) can be identified. Thus, to study theeffect of agents on cellular proliferation disorders, for example, in aclinical trial, cells can be isolated and RNA prepared and analyzed forthe levels of expression of a gene of the invention and other genesimplicated in the disorder. The levels of gene expression (i.e., a geneexpression pattern) can be quantified by Northern blot analysis orRT-PCR, as described herein, or alternatively by measuring the amount ofprotein produced, by one of the methods as described herein, or bymeasuring the levels of activity of a gene of the invention or othergenes. In this way, the gene expression pattern can serve as a marker,indicative of the physiological response of the cells to the agent.Accordingly, this response state can be determined before, and atvarious points during, treatment of the individual with the agent.

[0533] In a preferred embodiment, the present invention provides amethod for monitoring the effectiveness of treatment of a subject withan agent (e.g., an agonist, antagonist, peptidomimetic, protein,peptide, nucleic acid, small molecule, or other drug candidateidentified by the screening assays described herein) comprising thesteps of (i) obtaining a pre-administration sample from a subject priorto administration of the agent; (ii) detecting the level of thepolypeptide or nucleic acid of the invention in the pre-administrationsample; (iii) obtaining one or more post-administration samples from thesubject; (iv) detecting the level the of the polypeptide or nucleic acidof the invention in the post-administration samples; (v) comparing thelevel of the polypeptide or nucleic acid of the invention in thepre-administration sample with the level of the polypeptide or nucleicacid of the invention in the post-administration sample or samples; and(vi) altering the administration of the agent to the subjectaccordingly. For example, increased administration of the agent can bedesirable to increase the expression or activity of the polypeptide tohigher levels than detected, i.e., to increase the effectiveness of theagent. Alternatively, decreased administration of the agent can bedesirable to decrease expression or activity of the polypeptide to lowerlevels than detected, i.e., to decrease the effectiveness of the agent.

[0534] Methods of Treatment

[0535] The present invention provides for both prophylactic andtherapeutic methods of treating a subject at risk of (or susceptible to)a disorder or having a disorder associated with aberrant expression oractivity of a polypeptide of the invention and/or in which thepolypeptide of the invention is involved. Disorders characterized byaberrant expression or activity of the polypeptides of the invention aredescribed elsewhere in this disclosure.

[0536] Prophylactic Methods

[0537] In one aspect, the invention provides a method for preventing ina subject, a disease or condition associated with an aberrant expressionor activity of a polypeptide of the invention, by administering to thesubject an agent which modulates expression or at least one activity ofthe polypeptide. Subjects at risk for a disease which is caused orcontributed to by aberrant expression or activity of a polypeptide ofthe invention can be identified by, for example, any or a combination ofdiagnostic or prognostic assays as described herein. Administration of aprophylactic agent can occur prior to the manifestation of symptomscharacteristic of the aberrance, such that a disease or disorder isprevented or, alternatively, delayed in its progression. Depending onthe type of aberrance, for example, an agonist or antagonist agent canbe used for treating the subject. The appropriate agent can bedetermined based on screening assays described herein.

[0538] Therapeutic Methods

[0539] Another aspect of the invention pertains to methods of modulatingexpression or activity of a polypeptide of the invention for therapeuticpurposes. The modulatory method of the invention involves contacting acell with an agent that modulates one or more of the activities of thepolypeptide. An agent that modulates activity can be an agent asdescribed herein, such as a nucleic acid or a protein, anaturally-occurring cognate ligand of the polypeptide, a peptide, apeptidomimetic, or other small molecule. In one embodiment, the agentstimulates one or more of the biological activities of the polypeptide.Examples of such stimulatory agents include the active polypeptide ofthe invention and a nucleic acid molecule encoding the polypeptide ofthe invention that has been introduced into the cell. In anotherembodiment, the agent inhibits one or more of the biological activitiesof the polypeptide of the invention. Examples of such inhibitory agentsinclude antisense nucleic acid molecules and antibodies. Thesemodulatory methods can be performed in vitro (e.g., by culturing thecell with the agent) or, alternatively, in vivo (e.g., by administeringthe agent to a subject). As such, the present invention provides methodsof treating an individual afflicted with a disease or disordercharacterized by aberrant expression or activity of a polypeptide of theinvention. In one embodiment, the method involves administering an agent(e.g., an agent identified by a screening assay described herein), orcombination of agents that modulates (e.g., up-regulates ordown-regulates) expression or activity. In another embodiment, themethod involves administering a polypeptide of the invention or anucleic acid molecule of the invention as therapy to compensate forreduced or aberrant expression or activity of the polypeptide.

[0540] Stimulation of activity is desirable in situations in whichactivity or expression is abnormally low or down-regulated and/or inwhich increased activity is likely to have a beneficial effect.Conversely, inhibition of activity is desirable in situations in whichactivity or expression is abnormally high or up-regulated and/or inwhich decreased activity is likely to have a beneficial effect.

[0541] The contents of all references, patents, and published patentapplications cited throughout this application are hereby incorporatedby reference.

[0542] Deposit of Clones

[0543] Each of these deposits was made merely as a convenience to thoseof skill in the art. These deposits are not an admission that a depositis required under 35 U.S.C. §112.

[0544] Clone EpT202, encoding human TANGO 202 was deposited with theAmerican Type Culture Collection (ATCC®, 10801 University Boulevard,Manassas, V. 20110-2209) on Apr. 21, 1999 and was assigned AccessionNumber 207219. This deposit will be maintained under the terms of theBudapest Treaty on the International Recognition of the Deposit ofMicroorganisms for the Purposes of Patent Procedure. Clone EpTm202,encoding murine TANGO 202 was deposited with ATCC® on Apr. 21, 1999 andwas assigned (composite) Accession Number 207221. This deposit will bemaintained under the terms of the Budapest Treaty on the InternationalRecognition of the Deposit of Microorganisms for the Purposes of PatentProcedure.

[0545] Clone EpT234, encoding human TANGO 234 was deposited with ATCC®on Apr. 2, 1999 and was assigned Accession Number 207184. This depositwill be maintained under the terms of the Budapest Treaty on theInternational Recognition of the Deposit of Microorganisms for thePurposes of Patent Procedure.

[0546] Clone EpT265, encoding human TANGO 265 was deposited with ATCC®on Apr. 28, 1999 and was assigned Accession Number 207228. This depositwill be maintained under the terms of the Budapest Treaty on theInternational Recognition of the Deposit of Microorganisms for thePurposes of Patent Procedure.

[0547] Clone EpT273, encoding human TANGO 273 was deposited with ATCC®on Apr. 2, 1999 and was assigned Accession Number 207185. This depositwill be maintained under the terms of the Budapest Treaty on theInternational Recognition of the Deposit of Microorganisms for thePurposes of Patent Procedure.

[0548] Clone EpTm273, encoding murine TANGO 273 was deposited with ATCC®on Apr. 2, 1999 and was assigned (composite) Accession Number 207221.This deposit will be maintained under the terms of the Budapest Treatyon the International Recognition of the Deposit of Microorganisms forthe Purposes of Patent Procedure.

[0549] Clone EpT286, encoding human TANGO 286 was deposited with ATCC®on Apr. 20, 1999 and was assigned (composite) Accession Number 207220.This deposit will be maintained under the terms of the Budapest Treatyon the International Recognition of the Deposit of Microorganisms forthe Purposes of Patent Procedure.

[0550] Clone EpT294, encoding human TANGO 294 was deposited with ATCC®on Apr. 20, 1999 and was assigned (composite) Accession Number 207220.This deposit will be maintained under the terms of the Budapest Treatyon the International Recognition of the Deposit of Microorganisms forthe Purposes of Patent Procedure.

[0551] Clone EpT296, encoding human INTERCEPT 296 was deposited withATCC® on Apr. 20, 1999 and was assigned (composite) Accession Number207220. This deposit will be maintained under the terms of the BudapestTreaty on the International Recognition of the Deposit of Microorganismsfor the Purposes of Patent Procedure.

[0552] Clones containing cDNA molecules encoding human TANGO 286, humanTANGO 294, and INTERCEPT 296 were deposited with ATCC® on Apr. 21, 1999as Accession Number 207220, as part of a composite deposit representinga mixture of five strains, each carrying one recombinant plasmidharboring a particular cDNA clone.

[0553] To distinguish the strains and isolate a strain harboring aparticular cDNA clone, an aliquot of the mixture is streaked out tosingle colonies on nutrient medium (e.g., LB plates) supplemented with100 mg/ml ampicillin, single colonies are grown, and then plasmid DNA isextracted using a standard mini-preparation procedure. Next, a sample ofthe DNA mini-preparation is digested with a combination of therestriction enzymes SalI, NotI, and DraII and the resulting products areresolved on a 0.8% agarose gel using standard DNA electrophoresisconditions. This digestion procedure liberates fragments as follows:

[0554] 1. human TANGO 286 (clone EpT286): 1.85 kB and 0.1 kB (humanTANGO 286 has a DraII cut site at about base pair 1856).

[0555] 2. human TANGO 294 (clone EpT294): 1.4 kB and 0.6 kB (human TANGO294 has a DraII cut site at about base pair 1447).

[0556] 3. human INTERCEPT 296 (clone EpT296): 0.4 kB, 1.6 kB, and 0.1 kB(human INTERCEPT 296 has DraII cut sites at about base pair 410 and atabout base pair 1933). The identity of the strains can be inferred fromthe fragments liberated.

[0557] Clones containing cDNA molecules encoding mouse TANGO 202 andmouse TANGO 273 were deposited with ATCC® on Apr. 21, 1999 and wereassigned Accession Number 207221, as part of a composite depositrepresenting a mixture of five strains, each carrying one recombinantplasmid harboring a particular cDNA clone. To distinguish the strainsand isolate a strain harboring a particular cDNA clone, an aliquot ofthe mixture is streaked out to single colonies on nutrient medium (e.g.,LB plates) supplemented with 100 mg/ml ampicillin, single colonies aregrown, and then plasmid DNA is extracted using a standardmini-preparation procedure. Next, a sample of the DNA mini-preparationis digested with a combination of the restriction enzymes Sal I, Not I,and Apa I, and the resultant products are resolved on a 0.8% agarose gelusing standard DNA electrophoresis conditions. This digestion procedureliberates fragments as follows:

[0558] 1. mouse TANGO 202 (clone EpTm202): 3.5 kB and 1.4 kB (mouseTANGO 202 has a Apa I cut site at about base pair 3519).

[0559] 2. mouse TANGO 273 (clone EpTm273): 0.3 kB and 2.6 kB (mouseTANGO 273 has a Apa I cut site at about base pair 298).

[0560] The identity of the strains can be inferred from the fragmentsliberated.

[0561] Human TANGO 202, human TANGO 234, human TANGO 265, and humanTANGO 273 were each deposited as single deposits. Their clone names,deposit dates, and accession numbers are as follows:

[0562] 1. human TANGO 202: clone EpT202 was deposited with ATCC® on Apr.21, 1999, and was assigned Accession Number 207219.

[0563] 2. human TANGO 234: clone EpT234 was deposited with ATCC® on Apr.2, 1999, and was assigned Accession Number 207184.

[0564] 3. human TANGO 265: clone EpT265 was deposited with ATCC® on Apr.28, 1999, and was assigned Accession Number 207228.

[0565] 4. human TANGO 273: clone EpT273 was deposited with ATCC® on Apr.2, 1999, and was assigned Accession Number 207185.

[0566] All publications, patents, and patent applications referenced inthis specification are incorporated by reference into the specificationto the same extent as if each individual publication, patent, or patentapplication had been specifically and individually indicated to beincorporated herein by reference.

[0567] Equivalents

[0568] Those skilled in the art will recognize, or be able to ascertainusing no more than routine experimentation, many equivalents to thespecific embodiments of the invention described herein. Such equivalentsare intended to be encompassed by the following claims.

1 79 1 1656 DNA Homo sapiens 1 gtcgacccac gcgtccgccc acgcgtccggcccatggcgc cgcccgccgc ccgcctcgcc 60 ctgctctccg ccgcggcgct cacgctggcggcccggcccg cgcctagccc cggcctcggc 120 cccggacccg agtgtttcac agccaatggtgcggattata ggggaacaca gaactggaca 180 gcactacaag gcgggaagcc atgtctgttttggaacgaga ctttccagca tccatacaac 240 actctgaaat accccaacgg ggaggggggcctgggtgagc acaactattg cagaaatcca 300 gatggagacg tgagcccctg gtgctatgtggcagagcacg aggatggtgt ctactggaag 360 tactgtgaga tacctgcttg ccagatgcctggaaaccttg gctgctacaa ggatcatgga 420 aacccacctc ctctaactgg caccagtaaaacgtccaaca aactcaccat acaaacttgc 480 atcagttttt gtcggagtca gaggttcaagtttgctggga tggagtcagg ctatgcttgc 540 ttctgtggaa acaatcctga ttactggaagtacggggagg cagccagtac cgaatgcaac 600 agcgtctgct tcggggatca cacccaaccctgtggtggcg atggcaggat catcctcttt 660 gatactctcg tgggcgcctg cggtgggaactactcagcca tgtcttctgt ggtctattcc 720 cctgacttcc ccgacaccta tgccacggggagggtctgct actggaccat ccgggttccg 780 ggggcctccc acatccactt cagcttccccctatttgaca tcagggactc ggcggacatg 840 gtggagcttc tggatggcta cacccaccgtgtcctagccc gcttccacgg gaggagccgc 900 ccacctctgt ccttcaacgt ctctctggacttcgtcatct tgtatttctt ctctgatcgc 960 atcaatcagg cccagggatt tgctgttttataccaagccg tcaaggaaga actgccacag 1020 gagaggcccg ctgtcaacca gacggtggccgaggtgatca cggagcaggc caacctcagt 1080 gtcagcgctg cccggtcctc caaagtcctctatgtcatca ccaccagccc cagccaccca 1140 cctcagactg tcccaggtag caattcctgggcgccaccca tgggggctgg aagccacaga 1200 gttgaaggat ggacagtcta tggtctggcaactctcctca tcctcacagt cacagccatt 1260 gtagcaaaga tacttctgca cgtcacattcaaatcccatc gtgttcctgc ttcaggggac 1320 cttagggatt gtcatcaacc agggacttcgggggaaatct ggagcatttt ttacaagcct 1380 tccacttcaa tttccatctt taagaagaaactcaagggtc agagtcaaca agatgaccgc 1440 aatccccttg tgagtgacta aaaaccccactgtgcctagg acttgaggtc cctctttgag 1500 ctcaaggctg ccgtggtcaa cctctcctgtggttcttctc tgacagactc ttccctcctc 1560 tccctctgcc tcggcctctt cggggaaaccctcctcctac agactaggaa gaggcacctg 1620 ctgccagggc aggcagagcc tggattcctcctgctt 1656 2 1425 DNA Homo sapiens 2 atggcgccgc ccgccgcccg cctcgccctgctctccgccg cggcgctcac gctggcggcc 60 cggcccgcgc ctagccccgg cctcggccccggacccgagt gtttcacagc caatggtgcg 120 gattataggg gaacacagaa ctggacagcactacaaggcg ggaagccatg tctgttttgg 180 aacgagactt tccagcatcc atacaacactctgaaatacc ccaacgggga ggggggcctg 240 ggtgagcaca actattgcag aaatccagatggagacgtga gcccctggtg ctatgtggca 300 gagcacgagg atggtgtcta ctggaagtactgtgagatac ctgcttgcca gatgcctgga 360 aaccttggct gctacaagga tcatggaaacccacctcctc taactggcac cagtaaaacg 420 tccaacaaac tcaccataca aacttgcatcagtttttgtc ggagtcagag gttcaagttt 480 gctgggatgg agtcaggcta tgcttgcttctgtggaaaca atcctgatta ctggaagtac 540 ggggaggcag ccagtaccga atgcaacagcgtctgcttcg gggatcacac ccaaccctgt 600 ggtggcgatg gcaggatcat cctctttgatactctcgtgg gcgcctgcgg tgggaactac 660 tcagccatgt cttctgtggt ctattcccctgacttccccg acacctatgc cacggggagg 720 gtctgctact ggaccatccg ggttccgggggcctcccaca tccacttcag cttcccccta 780 tttgacatca gggactcggc ggacatggtggagcttctgg atggctacac ccaccgtgtc 840 ctagcccgct tccacgggag gagccgcccacctctgtcct tcaacgtctc tctggacttc 900 gtcatcttgt atttcttctc tgatcgcatcaatcaggccc agggatttgc tgttttatac 960 caagccgtca aggaagaact gccacaggagaggcccgctg tcaaccagac ggtggccgag 1020 gtgatcacgg agcaggccaa cctcagtgtcagcgctgccc ggtcctccaa agtcctctat 1080 gtcatcacca ccagccccag ccacccacctcagactgtcc caggtagcaa ttcctgggcg 1140 ccacccatgg gggctggaag ccacagagttgaaggatgga cagtctatgg tctggcaact 1200 ctcctcatcc tcacagtcac agccattgtagcaaagatac ttctgcacgt cacattcaaa 1260 tcccatcgtg ttcctgcttc aggggaccttagggattgtc atcaaccagg gacttcgggg 1320 gaaatctgga gcatttttta caagccttccacttcaattt ccatctttaa gaagaaactc 1380 aagggtcaga gtcaacaaga tgaccgcaatccccttgtga gtgac 1425 3 475 PRT Homo sapiens 3 Met Ala Pro Pro Ala AlaArg Leu Ala Leu Leu Ser Ala Ala Ala Leu 1 5 10 15 Thr Leu Ala Ala ArgPro Ala Pro Ser Pro Gly Leu Gly Pro Gly Pro 20 25 30 Glu Cys Phe Thr AlaAsn Gly Ala Asp Tyr Arg Gly Thr Gln Asn Trp 35 40 45 Thr Ala Leu Gln GlyGly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe 50 55 60 Gln His Pro Tyr AsnThr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu 65 70 75 80 Gly Glu His AsnTyr Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp 85 90 95 Cys Tyr Val AlaGlu His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu 100 105 110 Ile Pro AlaCys Gln Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His 115 120 125 Gly AsnPro Pro Pro Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu 130 135 140 ThrIle Gln Thr Cys Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe 145 150 155160 Ala Gly Met Glu Ser Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro Asp 165170 175 Tyr Trp Lys Tyr Gly Glu Ala Ala Ser Thr Glu Cys Asn Ser Val Cys180 185 190 Phe Gly Asp His Thr Gln Pro Cys Gly Gly Asp Gly Arg Ile IleLeu 195 200 205 Phe Asp Thr Leu Val Gly Ala Cys Gly Gly Asn Tyr Ser AlaMet Ser 210 215 220 Ser Val Val Tyr Ser Pro Asp Phe Pro Asp Thr Tyr AlaThr Gly Arg 225 230 235 240 Val Cys Tyr Trp Thr Ile Arg Val Pro Gly AlaSer His Ile His Phe 245 250 255 Ser Phe Pro Leu Phe Asp Ile Arg Asp SerAla Asp Met Val Glu Leu 260 265 270 Leu Asp Gly Tyr Thr His Arg Val LeuAla Arg Phe His Gly Arg Ser 275 280 285 Arg Pro Pro Leu Ser Phe Asn ValSer Leu Asp Phe Val Ile Leu Tyr 290 295 300 Phe Phe Ser Asp Arg Ile AsnGln Ala Gln Gly Phe Ala Val Leu Tyr 305 310 315 320 Gln Ala Val Lys GluGlu Leu Pro Gln Glu Arg Pro Ala Val Asn Gln 325 330 335 Thr Val Ala GluVal Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala 340 345 350 Ala Arg SerSer Lys Val Leu Tyr Val Ile Thr Thr Ser Pro Ser His 355 360 365 Pro ProGln Thr Val Pro Gly Ser Asn Ser Trp Ala Pro Pro Met Gly 370 375 380 AlaGly Ser His Arg Val Glu Gly Trp Thr Val Tyr Gly Leu Ala Thr 385 390 395400 Leu Leu Ile Leu Thr Val Thr Ala Ile Val Ala Lys Ile Leu Leu His 405410 415 Val Thr Phe Lys Ser His Arg Val Pro Ala Ser Gly Asp Leu Arg Asp420 425 430 Cys His Gln Pro Gly Thr Ser Gly Glu Ile Trp Ser Ile Phe TyrLys 435 440 445 Pro Ser Thr Ser Ile Ser Ile Phe Lys Lys Lys Leu Lys GlyGln Ser 450 455 460 Gln Gln Asp Asp Arg Asn Pro Leu Val Ser Asp 465 470475 4 19 PRT Homo sapiens 4 Met Ala Pro Pro Ala Ala Arg Leu Ala Leu LeuSer Ala Ala Ala Leu 1 5 10 15 Thr Leu Ala 5 456 PRT Homo sapiens 5 AlaArg Pro Ala Pro Ser Pro Gly Leu Gly Pro Gly Pro Glu Cys Phe 1 5 10 15Thr Ala Asn Gly Ala Asp Tyr Arg Gly Thr Gln Asn Trp Thr Ala Leu 20 25 30Gln Gly Gly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe Gln His Pro 35 40 45Tyr Asn Thr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu Gly Glu His 50 55 60Asn Tyr Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp Cys Tyr Val 65 70 7580 Ala Glu His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu Ile Pro Ala 85 9095 Cys Gln Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His Gly Asn Pro 100105 110 Pro Pro Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu Thr Ile Gln115 120 125 Thr Cys Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe Ala GlyMet 130 135 140 Glu Ser Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro Asp TyrTrp Lys 145 150 155 160 Tyr Gly Glu Ala Ala Ser Thr Glu Cys Asn Ser ValCys Phe Gly Asp 165 170 175 His Thr Gln Pro Cys Gly Gly Asp Gly Arg IleIle Leu Phe Asp Thr 180 185 190 Leu Val Gly Ala Cys Gly Gly Asn Tyr SerAla Met Ser Ser Val Val 195 200 205 Tyr Ser Pro Asp Phe Pro Asp Thr TyrAla Thr Gly Arg Val Cys Tyr 210 215 220 Trp Thr Ile Arg Val Pro Gly AlaSer His Ile His Phe Ser Phe Pro 225 230 235 240 Leu Phe Asp Ile Arg AspSer Ala Asp Met Val Glu Leu Leu Asp Gly 245 250 255 Tyr Thr His Arg ValLeu Ala Arg Phe His Gly Arg Ser Arg Pro Pro 260 265 270 Leu Ser Phe AsnVal Ser Leu Asp Phe Val Ile Leu Tyr Phe Phe Ser 275 280 285 Asp Arg IleAsn Gln Ala Gln Gly Phe Ala Val Leu Tyr Gln Ala Val 290 295 300 Lys GluGlu Leu Pro Gln Glu Arg Pro Ala Val Asn Gln Thr Val Ala 305 310 315 320Glu Val Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala Ala Arg Ser 325 330335 Ser Lys Val Leu Tyr Val Ile Thr Thr Ser Pro Ser His Pro Pro Gln 340345 350 Thr Val Pro Gly Ser Asn Ser Trp Ala Pro Pro Met Gly Ala Gly Ser355 360 365 His Arg Val Glu Gly Trp Thr Val Tyr Gly Leu Ala Thr Leu LeuIle 370 375 380 Leu Thr Val Thr Ala Ile Val Ala Lys Ile Leu Leu His ValThr Phe 385 390 395 400 Lys Ser His Arg Val Pro Ala Ser Gly Asp Leu ArgAsp Cys His Gln 405 410 415 Pro Gly Thr Ser Gly Glu Ile Trp Ser Ile PheTyr Lys Pro Ser Thr 420 425 430 Ser Ile Ser Ile Phe Lys Lys Lys Leu LysGly Gln Ser Gln Gln Asp 435 440 445 Asp Arg Asn Pro Leu Val Ser Asp 450455 6 373 PRT Homo sapiens 6 Ala Arg Pro Ala Pro Ser Pro Gly Leu Gly ProGly Pro Glu Cys Phe 1 5 10 15 Thr Ala Asn Gly Ala Asp Tyr Arg Gly ThrGln Asn Trp Thr Ala Leu 20 25 30 Gln Gly Gly Lys Pro Cys Leu Phe Trp AsnGlu Thr Phe Gln His Pro 35 40 45 Tyr Asn Thr Leu Lys Tyr Pro Asn Gly GluGly Gly Leu Gly Glu His 50 55 60 Asn Tyr Cys Arg Asn Pro Asp Gly Asp ValSer Pro Trp Cys Tyr Val 65 70 75 80 Ala Glu His Glu Asp Gly Val Tyr TrpLys Tyr Cys Glu Ile Pro Ala 85 90 95 Cys Gln Met Pro Gly Asn Leu Gly CysTyr Lys Asp His Gly Asn Pro 100 105 110 Pro Pro Leu Thr Gly Thr Ser LysThr Ser Asn Lys Leu Thr Ile Gln 115 120 125 Thr Cys Ile Ser Phe Cys ArgSer Gln Arg Phe Lys Phe Ala Gly Met 130 135 140 Glu Ser Gly Tyr Ala CysPhe Cys Gly Asn Asn Pro Asp Tyr Trp Lys 145 150 155 160 Tyr Gly Glu AlaAla Ser Thr Glu Cys Asn Ser Val Cys Phe Gly Asp 165 170 175 His Thr GlnPro Cys Gly Gly Asp Gly Arg Ile Ile Leu Phe Asp Thr 180 185 190 Leu ValGly Ala Cys Gly Gly Asn Tyr Ser Ala Met Ser Ser Val Val 195 200 205 TyrSer Pro Asp Phe Pro Asp Thr Tyr Ala Thr Gly Arg Val Cys Tyr 210 215 220Trp Thr Ile Arg Val Pro Gly Ala Ser His Ile His Phe Ser Phe Pro 225 230235 240 Leu Phe Asp Ile Arg Asp Ser Ala Asp Met Val Glu Leu Leu Asp Gly245 250 255 Tyr Thr His Arg Val Leu Ala Arg Phe His Gly Arg Ser Arg ProPro 260 265 270 Leu Ser Phe Asn Val Ser Leu Asp Phe Val Ile Leu Tyr PhePhe Ser 275 280 285 Asp Arg Ile Asn Gln Ala Gln Gly Phe Ala Val Leu TyrGln Ala Val 290 295 300 Lys Glu Glu Leu Pro Gln Glu Arg Pro Ala Val AsnGln Thr Val Ala 305 310 315 320 Glu Val Ile Thr Glu Gln Ala Asn Leu SerVal Ser Ala Ala Arg Ser 325 330 335 Ser Lys Val Leu Tyr Val Ile Thr ThrSer Pro Ser His Pro Pro Gln 340 345 350 Thr Val Pro Gly Ser Asn Ser TrpAla Pro Pro Met Gly Ala Gly Ser 355 360 365 His Arg Val Glu Gly 370 7 23PRT Homo sapiens 7 Trp Thr Val Tyr Gly Leu Ala Thr Leu Leu Ile Leu ThrVal Thr Ala 1 5 10 15 Ile Val Ala Lys Ile Leu Leu 20 8 60 PRT Homosapiens 8 His Val Thr Phe Lys Ser His Arg Val Pro Ala Ser Gly Asp LeuArg 1 5 10 15 Asp Cys His Gln Pro Gly Thr Ser Gly Glu Ile Trp Ser IlePhe Tyr 20 25 30 Lys Pro Ser Thr Ser Ile Ser Ile Phe Lys Lys Lys Leu LysGly Gln 35 40 45 Ser Gln Gln Asp Asp Arg Asn Pro Leu Val Ser Asp 50 5560 9 4628 DNA Homo sapiens 9 gcggccgctc gcgatctaga actagtaatg atgctgcctcaaaactcgtg gcatattgat 60 tttggaagat gctgctgtca tcagaacctt ttctctgctgtggtaacttg catcctgctc 120 ctgaattcct gctttctcat cagcagtttt aatggaacagatttggagtt gaggctggtc 180 aatggagacg gtccctgctc tgggacagtg gaggtgaaattccagggaca gtgggggact 240 gtgtgtgatg atgggtggaa cactactgcc tcaactgtcgtgtgcaaaca gcttggatgt 300 ccattttctt tcgccatgtt tcgttttgga caagccgtgactagacatgg aaaaatttgg 360 cttgatgatg tttcctgtta tggaaatgag tcagctctctgggaatgtca acaccgggaa 420 tggggaagcc ataactgtta tcatggagaa gatgttggtgtgaactgtta tggtgaagcc 480 aatctgggtt tgaggctagt ggatggaaac aactcctgttcagggagagt ggaggtgaaa 540 ttccaagaaa ggtgggggac tatatgtgat gatgggtggaacttgaatac tgctgccgtg 600 gtgtgcaggc aactaggatg tccatcttct tttatttcttctggagttgt taatagccct 660 gctgtattgc gccccatttg gctggatgac attttatgccaggggaatga gttggcactc 720 tggaattgca gacatcgtgg atggggaaat catgactgcagtcacaatga ggatgtcaca 780 ttaacttgtt atgatagtag tgatcttgaa ctaaggcttgtaggtggaac taaccgctgt 840 atggggagag tagagctgaa aatccaagga aggtgggggaccgtatgcca ccataagtgg 900 aacaatgctg cagctgatgt cgtatgcaag cagttgggatgtggaaccgc acttcacttc 960 gctggcttgc ctcatttgca gtcagggtct gatgttgtatggcttgatgg tgtctcctgc 1020 tccggtaatg aatcttttct ttgggactgc agacattccggaaccgtcaa ttttgactgt 1080 cttcatcaaa acgatgtgtc tgtgatctgc tcagatggagcagatttgga actgcgacta 1140 gcagatggaa gtaacaattg ttcagggaga gtagaggtgagaattcatga acagtggtgg 1200 acaatatgtg accagaactg gaagaatgaa caagcccttgtggtttgtaa gcagctagga 1260 tgtccgttca gcgtctttgg cagtcgtcgt gctaaacctagtaatgaagc tagagacatt 1320 tggataaaca gcatatcttg cactgggaat gagtcagctctctgggactg cacatatgat 1380 ggaaaagcaa agcgaacatg cttccgaaga tcagatgctggagtaatttg ttctgataag 1440 gcagatctgg acctaaggct tgtcggggct catagcccctgttatgggag attggaggtg 1500 aaataccaag gagagtgggg gactgtgtgt catgacagatggagcacaag gaatgcagct 1560 gttgtgtgta aacaattggg atgtggaaag cctatgcatgtgtttggtat gacctatttt 1620 aaagaagcat caggacctat ttggctggat gacgtttcttgcattggaaa tgagtcaaat 1680 atctgggact gtgaacacag tggatgggga aagcataattgtgtacacag agaggatgtg 1740 attgtaacct gctcaggtga tgcaacatgg ggcctgaggctggtgggcgg cagcaaccgc 1800 tgctcgggaa gactggaggt gtactttcaa ggacggtggggcacagtgtg tgatgacggc 1860 tggaacagta aagctgcagc tgtggtgtgt agccagctggactgcccatc ttctatcatt 1920 ggcatgggtc tgggaaacgc ttctacagga tatggaaaaatttggctcga tgatgtttcc 1980 tgtgatggag atgagtcaga tctctggtca tgcaggaacagtgggtgggg aaataatgac 2040 tgcagtcaca gtgaagatgt tggagtgatc tgttctgatgcatcggatat ggagctgagg 2100 cttgtgggtg gaagcagcag gtgtgctgga aaagttgaggtgaatgtcca gggtgccgtg 2160 ggaattctgt gtgctaatgg ctggggaatg aacattgctgaagttgtttg caggcaactt 2220 gaatgtgggt ctgcaatcag ggtctccaga gagcctcatttcacagaaag aacattacac 2280 atcttaatgt cgaattctgg ctgcactgga ggggaagcctctctctggga ttgtatacga 2340 tgggagtgga aacagactgc gtgtcattta aatatggaagcaagtttgat ctgctcagcc 2400 cacaggcagc ccaggctggt tggagctgat atgccctgctctggacgtgt tgaagtgaaa 2460 catgcagaca catggcgctc tgtctgtgat tctgatttctctcttcatgc tgccaatgtg 2520 ctgtgcagag aattaaattg tggagatgcc atatctctttctgtgggaga tcactttgga 2580 aaagggaatg gtctaacttg ggccgaaaag ttccagtgtgaagggagtga aactcacctt 2640 gcattatgcc ccattgttca acatccggaa gacacttgtatccacagcag agaagttgga 2700 gttgtctgtt cccgatatac agatgtccga cttgtgaatggcaaatccca gtgtgacggg 2760 caagtggaga tcaacgtgct tggacactgg ggctcactgtgtgacaccca ctgggaccca 2820 gaagatgccc gtgttctatg cagacagctc agctgtgggactgctctctc aaccacagga 2880 ggaaaatata ttggagaaag aagtgttcgt gtgtggggacacaggtttca ttgcttaggg 2940 aatgagtcac ttctggataa ctgtcaaatg acagttcttggagcacctcc ctgtatccat 3000 ggaaatactg tctctgtgat ctgcacagga agcctgacccagccactgtt tccatgcctc 3060 gcaaatgtat ctgacccata tttgtctgca gttccagagggcagtgcttt gatctgctta 3120 gaggacaaac ggctccgcct agtggatggg gacagccgctgtgccgggag agtagagatc 3180 tatcacgacg gcttctgggg caccatctgt gatgacggctgggacctgag cgatgcccac 3240 gtggtgtgtc aaaagctggg ctgtggagtg gccttcaatgccacggtctc tgctcacttt 3300 ggggaggggt cagggcccat ctggctggat gacctgaactgcacaggaac ggagtcccac 3360 ttgtggcagt gcccttcccg cggctggggg cagcacgactgcaggcacaa ggaggacgca 3420 ggggtcatct gctcagaatt cacagccttg aggctctacagtgaaactga aacagagagc 3480 tgtgctggga gattggaagt cttctataac gggacctggggcagcgtcgg caggaggaac 3540 atcaccacag ccatagcagg cattgtgtgc aggcagctgggctgtgggga gaatggagtt 3600 gtcagcctcg cccctttatc taagacaggc tctggtttcatgtgggtgga tgacattcag 3660 tgtcctaaaa cgcatatctc catatggcag tgcctgtctgccccatggga gcgaagaatc 3720 tccagcccag cagaagagac ctggatcaca tgtgaagatagaataagagt gcgtggagga 3780 gacaccgagt gctctgggag agtggagatc tggcacgcaggctcctgggg cacagtgtgt 3840 gatgactcct gggacctggc cgaggcggaa gtggtgtgtcagcagctggg ctgtggctct 3900 gctctggctg ccctgaggga cgcttcgttt ggccagggaactggaaccat ctggttggat 3960 gacatgcggt gcaaaggaaa tgagtcattt ctatgggactgtcacgccaa accctgggga 4020 cagagtgact gtggacacaa ggaagatgct ggcgtgaggtgctctggaca gtcgctgaaa 4080 tcactgaatg cctcctcagg tcatttagca cttattttatccagtatctt tgggctcctt 4140 ctcctggttc tgtttattct atttctcacg tggtgccgagttcagaaaca aaaacatctg 4200 cccctcagag tttcaaccag aaggaggggt tctctcgaggagaatttatt ccatgagatg 4260 gagacctgcc tcaagagaga ggacccacat gggacaagaacctcagatga cacccccaac 4320 catggttgtg aagatgctag cgacacatcg ctgttgggagttcttcctgc ctctgaagcc 4380 acaaaatgac tttagacttc cagggctcac cagatcaacctctaaatatc tttgaaggag 4440 acaacaactt ttaaatgaat aaagaggaag tcaagttgccctatggaaaa cttgtccaaa 4500 taacatttct tgaacaatag gagaacagct aaattgataaagactggtga taataaaaat 4560 tgaattatgt atatcactgt taaaaaaaaa aaaaaaaaaaaaaaaaaaaa acggacgcgt 4620 gggtcgac 4628 10 4359 DNA Homo sapiens 10atgatgctgc ctcaaaactc gtggcatatt gattttggaa gatgctgctg tcatcagaac 60cttttctctg ctgtggtaac ttgcatcctg ctcctgaatt cctgctttct catcagcagt 120tttaatggaa cagatttgga gttgaggctg gtcaatggag acggtccctg ctctgggaca 180gtggaggtga aattccaggg acagtggggg actgtgtgtg atgatgggtg gaacactact 240gcctcaactg tcgtgtgcaa acagcttgga tgtccatttt ctttcgccat gtttcgtttt 300ggacaagccg tgactagaca tggaaaaatt tggcttgatg atgtttcctg ttatggaaat 360gagtcagctc tctgggaatg tcaacaccgg gaatggggaa gccataactg ttatcatgga 420gaagatgttg gtgtgaactg ttatggtgaa gccaatctgg gtttgaggct agtggatgga 480aacaactcct gttcagggag agtggaggtg aaattccaag aaaggtgggg gactatatgt 540gatgatgggt ggaacttgaa tactgctgcc gtggtgtgca ggcaactagg atgtccatct 600tcttttattt cttctggagt tgttaatagc cctgctgtat tgcgccccat ttggctggat 660gacattttat gccaggggaa tgagttggca ctctggaatt gcagacatcg tggatgggga 720aatcatgact gcagtcacaa tgaggatgtc acattaactt gttatgatag tagtgatctt 780gaactaaggc ttgtaggtgg aactaaccgc tgtatgggga gagtagagct gaaaatccaa 840ggaaggtggg ggaccgtatg ccaccataag tggaacaatg ctgcagctga tgtcgtatgc 900aagcagttgg gatgtggaac cgcacttcac ttcgctggct tgcctcattt gcagtcaggg 960tctgatgttg tatggcttga tggtgtctcc tgctccggta atgaatcttt tctttgggac 1020tgcagacatt ccggaaccgt caattttgac tgtcttcatc aaaacgatgt gtctgtgatc 1080tgctcagatg gagcagattt ggaactgcga ctagcagatg gaagtaacaa ttgttcaggg 1140agagtagagg tgagaattca tgaacagtgg tggacaatat gtgaccagaa ctggaagaat 1200gaacaagccc ttgtggtttg taagcagcta ggatgtccgt tcagcgtctt tggcagtcgt 1260cgtgctaaac ctagtaatga agctagagac atttggataa acagcatatc ttgcactggg 1320aatgagtcag ctctctggga ctgcacatat gatggaaaag caaagcgaac atgcttccga 1380agatcagatg ctggagtaat ttgttctgat aaggcagatc tggacctaag gcttgtcggg 1440gctcatagcc cctgttatgg gagattggag gtgaaatacc aaggagagtg ggggactgtg 1500tgtcatgaca gatggagcac aaggaatgca gctgttgtgt gtaaacaatt gggatgtgga 1560aagcctatgc atgtgtttgg tatgacctat tttaaagaag catcaggacc tatttggctg 1620gatgacgttt cttgcattgg aaatgagtca aatatctggg actgtgaaca cagtggatgg 1680ggaaagcata attgtgtaca cagagaggat gtgattgtaa cctgctcagg tgatgcaaca 1740tggggcctga ggctggtggg cggcagcaac cgctgctcgg gaagactgga ggtgtacttt 1800caaggacggt ggggcacagt gtgtgatgac ggctggaaca gtaaagctgc agctgtggtg 1860tgtagccagc tggactgccc atcttctatc attggcatgg gtctgggaaa cgcttctaca 1920ggatatggaa aaatttggct cgatgatgtt tcctgtgatg gagatgagtc agatctctgg 1980tcatgcagga acagtgggtg gggaaataat gactgcagtc acagtgaaga tgttggagtg 2040atctgttctg atgcatcgga tatggagctg aggcttgtgg gtggaagcag caggtgtgct 2100ggaaaagttg aggtgaatgt ccagggtgcc gtgggaattc tgtgtgctaa tggctgggga 2160atgaacattg ctgaagttgt ttgcaggcaa cttgaatgtg ggtctgcaat cagggtctcc 2220agagagcctc atttcacaga aagaacatta cacatcttaa tgtcgaattc tggctgcact 2280ggaggggaag cctctctctg ggattgtata cgatgggagt ggaaacagac tgcgtgtcat 2340ttaaatatgg aagcaagttt gatctgctca gcccacaggc agcccaggct ggttggagct 2400gatatgccct gctctggacg tgttgaagtg aaacatgcag acacatggcg ctctgtctgt 2460gattctgatt tctctcttca tgctgccaat gtgctgtgca gagaattaaa ttgtggagat 2520gccatatctc tttctgtggg agatcacttt ggaaaaggga atggtctaac ttgggccgaa 2580aagttccagt gtgaagggag tgaaactcac cttgcattat gccccattgt tcaacatccg 2640gaagacactt gtatccacag cagagaagtt ggagttgtct gttcccgata tacagatgtc 2700cgacttgtga atggcaaatc ccagtgtgac gggcaagtgg agatcaacgt gcttggacac 2760tggggctcac tgtgtgacac ccactgggac ccagaagatg cccgtgttct atgcagacag 2820ctcagctgtg ggactgctct ctcaaccaca ggaggaaaat atattggaga aagaagtgtt 2880cgtgtgtggg gacacaggtt tcattgctta gggaatgagt cacttctgga taactgtcaa 2940atgacagttc ttggagcacc tccctgtatc catggaaata ctgtctctgt gatctgcaca 3000ggaagcctga cccagccact gtttccatgc ctcgcaaatg tatctgaccc atatttgtct 3060gcagttccag agggcagtgc tttgatctgc ttagaggaca aacggctccg cctagtggat 3120ggggacagcc gctgtgccgg gagagtagag atctatcacg acggcttctg gggcaccatc 3180tgtgatgacg gctgggacct gagcgatgcc cacgtggtgt gtcaaaagct gggctgtgga 3240gtggccttca atgccacggt ctctgctcac tttggggagg ggtcagggcc catctggctg 3300gatgacctga actgcacagg aacggagtcc cacttgtggc agtgcccttc ccgcggctgg 3360gggcagcacg actgcaggca caaggaggac gcaggggtca tctgctcaga attcacagcc 3420ttgaggctct acagtgaaac tgaaacagag agctgtgctg ggagattgga agtcttctat 3480aacgggacct ggggcagcgt cggcaggagg aacatcacca cagccatagc aggcattgtg 3540tgcaggcagc tgggctgtgg ggagaatgga gttgtcagcc tcgccccttt atctaagaca 3600ggctctggtt tcatgtgggt ggatgacatt cagtgtccta aaacgcatat ctccatatgg 3660cagtgcctgt ctgccccatg ggagcgaaga atctccagcc cagcagaaga gacctggatc 3720acatgtgaag atagaataag agtgcgtgga ggagacaccg agtgctctgg gagagtggag 3780atctggcacg caggctcctg gggcacagtg tgtgatgact cctgggacct ggccgaggcg 3840gaagtggtgt gtcagcagct gggctgtggc tctgctctgg ctgccctgag ggacgcttcg 3900tttggccagg gaactggaac catctggttg gatgacatgc ggtgcaaagg aaatgagtca 3960tttctatggg actgtcacgc caaaccctgg ggacagagtg actgtggaca caaggaagat 4020gctggcgtga ggtgctctgg acagtcgctg aaatcactga atgcctcctc aggtcattta 4080gcacttattt tatccagtat ctttgggctc cttctcctgg ttctgtttat tctatttctc 4140acgtggtgcc gagttcagaa acaaaaacat ctgcccctca gagtttcaac cagaaggagg 4200ggttctctcg aggagaattt attccatgag atggagacct gcctcaagag agaggaccca 4260catgggacaa gaacctcaga tgacaccccc aaccatggtt gtgaagatgc tagcgacaca 4320tcgctgttgg gagttcttcc tgcctctgaa gccacaaaa 4359 11 1453 PRT Homo sapiens11 Met Met Leu Pro Gln Asn Ser Trp His Ile Asp Phe Gly Arg Cys Cys 1 510 15 Cys His Gln Asn Leu Phe Ser Ala Val Val Thr Cys Ile Leu Leu Leu 2025 30 Asn Ser Cys Phe Leu Ile Ser Ser Phe Asn Gly Thr Asp Leu Glu Leu 3540 45 Arg Leu Val Asn Gly Asp Gly Pro Cys Ser Gly Thr Val Glu Val Lys 5055 60 Phe Gln Gly Gln Trp Gly Thr Val Cys Asp Asp Gly Trp Asn Thr Thr 6570 75 80 Ala Ser Thr Val Val Cys Lys Gln Leu Gly Cys Pro Phe Ser Phe Ala85 90 95 Met Phe Arg Phe Gly Gln Ala Val Thr Arg His Gly Lys Ile Trp Leu100 105 110 Asp Asp Val Ser Cys Tyr Gly Asn Glu Ser Ala Leu Trp Glu CysGln 115 120 125 His Arg Glu Trp Gly Ser His Asn Cys Tyr His Gly Glu AspVal Gly 130 135 140 Val Asn Cys Tyr Gly Glu Ala Asn Leu Gly Leu Arg LeuVal Asp Gly 145 150 155 160 Asn Asn Ser Cys Ser Gly Arg Val Glu Val LysPhe Gln Glu Arg Trp 165 170 175 Gly Thr Ile Cys Asp Asp Gly Trp Asn LeuAsn Thr Ala Ala Val Val 180 185 190 Cys Arg Gln Leu Gly Cys Pro Ser SerPhe Ile Ser Ser Gly Val Val 195 200 205 Asn Ser Pro Ala Val Leu Arg ProIle Trp Leu Asp Asp Ile Leu Cys 210 215 220 Gln Gly Asn Glu Leu Ala LeuTrp Asn Cys Arg His Arg Gly Trp Gly 225 230 235 240 Asn His Asp Cys SerHis Asn Glu Asp Val Thr Leu Thr Cys Tyr Asp 245 250 255 Ser Ser Asp LeuGlu Leu Arg Leu Val Gly Gly Thr Asn Arg Cys Met 260 265 270 Gly Arg ValGlu Leu Lys Ile Gln Gly Arg Trp Gly Thr Val Cys His 275 280 285 His LysTrp Asn Asn Ala Ala Ala Asp Val Val Cys Lys Gln Leu Gly 290 295 300 CysGly Thr Ala Leu His Phe Ala Gly Leu Pro His Leu Gln Ser Gly 305 310 315320 Ser Asp Val Val Trp Leu Asp Gly Val Ser Cys Ser Gly Asn Glu Ser 325330 335 Phe Leu Trp Asp Cys Arg His Ser Gly Thr Val Asn Phe Asp Cys Leu340 345 350 His Gln Asn Asp Val Ser Val Ile Cys Ser Asp Gly Ala Asp LeuGlu 355 360 365 Leu Arg Leu Ala Asp Gly Ser Asn Asn Cys Ser Gly Arg ValGlu Val 370 375 380 Arg Ile His Glu Gln Trp Trp Thr Ile Cys Asp Gln AsnTrp Lys Asn 385 390 395 400 Glu Gln Ala Leu Val Val Cys Lys Gln Leu GlyCys Pro Phe Ser Val 405 410 415 Phe Gly Ser Arg Arg Ala Lys Pro Ser AsnGlu Ala Arg Asp Ile Trp 420 425 430 Ile Asn Ser Ile Ser Cys Thr Gly AsnGlu Ser Ala Leu Trp Asp Cys 435 440 445 Thr Tyr Asp Gly Lys Ala Lys ArgThr Cys Phe Arg Arg Ser Asp Ala 450 455 460 Gly Val Ile Cys Ser Asp LysAla Asp Leu Asp Leu Arg Leu Val Gly 465 470 475 480 Ala His Ser Pro CysTyr Gly Arg Leu Glu Val Lys Tyr Gln Gly Glu 485 490 495 Trp Gly Thr ValCys His Asp Arg Trp Ser Thr Arg Asn Ala Ala Val 500 505 510 Val Cys LysGln Leu Gly Cys Gly Lys Pro Met His Val Phe Gly Met 515 520 525 Thr TyrPhe Lys Glu Ala Ser Gly Pro Ile Trp Leu Asp Asp Val Ser 530 535 540 CysIle Gly Asn Glu Ser Asn Ile Trp Asp Cys Glu His Ser Gly Trp 545 550 555560 Gly Lys His Asn Cys Val His Arg Glu Asp Val Ile Val Thr Cys Ser 565570 575 Gly Asp Ala Thr Trp Gly Leu Arg Leu Val Gly Gly Ser Asn Arg Cys580 585 590 Ser Gly Arg Leu Glu Val Tyr Phe Gln Gly Arg Trp Gly Thr ValCys 595 600 605 Asp Asp Gly Trp Asn Ser Lys Ala Ala Ala Val Val Cys SerGln Leu 610 615 620 Asp Cys Pro Ser Ser Ile Ile Gly Met Gly Leu Gly AsnAla Ser Thr 625 630 635 640 Gly Tyr Gly Lys Ile Trp Leu Asp Asp Val SerCys Asp Gly Asp Glu 645 650 655 Ser Asp Leu Trp Ser Cys Arg Asn Ser GlyTrp Gly Asn Asn Asp Cys 660 665 670 Ser His Ser Glu Asp Val Gly Val IleCys Ser Asp Ala Ser Asp Met 675 680 685 Glu Leu Arg Leu Val Gly Gly SerSer Arg Cys Ala Gly Lys Val Glu 690 695 700 Val Asn Val Gln Gly Ala ValGly Ile Leu Cys Ala Asn Gly Trp Gly 705 710 715 720 Met Asn Ile Ala GluVal Val Cys Arg Gln Leu Glu Cys Gly Ser Ala 725 730 735 Ile Arg Val SerArg Glu Pro His Phe Thr Glu Arg Thr Leu His Ile 740 745 750 Leu Met SerAsn Ser Gly Cys Thr Gly Gly Glu Ala Ser Leu Trp Asp 755 760 765 Cys IleArg Trp Glu Trp Lys Gln Thr Ala Cys His Leu Asn Met Glu 770 775 780 AlaSer Leu Ile Cys Ser Ala His Arg Gln Pro Arg Leu Val Gly Ala 785 790 795800 Asp Met Pro Cys Ser Gly Arg Val Glu Val Lys His Ala Asp Thr Trp 805810 815 Arg Ser Val Cys Asp Ser Asp Phe Ser Leu His Ala Ala Asn Val Leu820 825 830 Cys Arg Glu Leu Asn Cys Gly Asp Ala Ile Ser Leu Ser Val GlyAsp 835 840 845 His Phe Gly Lys Gly Asn Gly Leu Thr Trp Ala Glu Lys PheGln Cys 850 855 860 Glu Gly Ser Glu Thr His Leu Ala Leu Cys Pro Ile ValGln His Pro 865 870 875 880 Glu Asp Thr Cys Ile His Ser Arg Glu Val GlyVal Val Cys Ser Arg 885 890 895 Tyr Thr Asp Val Arg Leu Val Asn Gly LysSer Gln Cys Asp Gly Gln 900 905 910 Val Glu Ile Asn Val Leu Gly His TrpGly Ser Leu Cys Asp Thr His 915 920 925 Trp Asp Pro Glu Asp Ala Arg ValLeu Cys Arg Gln Leu Ser Cys Gly 930 935 940 Thr Ala Leu Ser Thr Thr GlyGly Lys Tyr Ile Gly Glu Arg Ser Val 945 950 955 960 Arg Val Trp Gly HisArg Phe His Cys Leu Gly Asn Glu Ser Leu Leu 965 970 975 Asp Asn Cys GlnMet Thr Val Leu Gly Ala Pro Pro Cys Ile His Gly 980 985 990 Asn Thr ValSer Val Ile Cys Thr Gly Ser Leu Thr Gln Pro Leu Phe 995 1000 1005 ProCys Leu Ala Asn Val Ser Asp Pro Tyr Leu Ser Ala Val Pro Glu 1010 10151020 Gly Ser Ala Leu Ile Cys Leu Glu Asp Lys Arg Leu Arg Leu Val Asp1025 1030 1035 1040 Gly Asp Ser Arg Cys Ala Gly Arg Val Glu Ile Tyr HisAsp Gly Phe 1045 1050 1055 Trp Gly Thr Ile Cys Asp Asp Gly Trp Asp LeuSer Asp Ala His Val 1060 1065 1070 Val Cys Gln Lys Leu Gly Cys Gly ValAla Phe Asn Ala Thr Val Ser 1075 1080 1085 Ala His Phe Gly Glu Gly SerGly Pro Ile Trp Leu Asp Asp Leu Asn 1090 1095 1100 Cys Thr Gly Thr GluSer His Leu Trp Gln Cys Pro Ser Arg Gly Trp 1105 1110 1115 1120 Gly GlnHis Asp Cys Arg His Lys Glu Asp Ala Gly Val Ile Cys Ser 1125 1130 1135Glu Phe Thr Ala Leu Arg Leu Tyr Ser Glu Thr Glu Thr Glu Ser Cys 11401145 1150 Ala Gly Arg Leu Glu Val Phe Tyr Asn Gly Thr Trp Gly Ser ValGly 1155 1160 1165 Arg Arg Asn Ile Thr Thr Ala Ile Ala Gly Ile Val CysArg Gln Leu 1170 1175 1180 Gly Cys Gly Glu Asn Gly Val Val Ser Leu AlaPro Leu Ser Lys Thr 1185 1190 1195 1200 Gly Ser Gly Phe Met Trp Val AspAsp Ile Gln Cys Pro Lys Thr His 1205 1210 1215 Ile Ser Ile Trp Gln CysLeu Ser Ala Pro Trp Glu Arg Arg Ile Ser 1220 1225 1230 Ser Pro Ala GluGlu Thr Trp Ile Thr Cys Glu Asp Arg Ile Arg Val 1235 1240 1245 Arg GlyGly Asp Thr Glu Cys Ser Gly Arg Val Glu Ile Trp His Ala 1250 1255 1260Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp Asp Leu Ala Glu Ala 12651270 1275 1280 Glu Val Val Cys Gln Gln Leu Gly Cys Gly Ser Ala Leu AlaAla Leu 1285 1290 1295 Arg Asp Ala Ser Phe Gly Gln Gly Thr Gly Thr IleTrp Leu Asp Asp 1300 1305 1310 Met Arg Cys Lys Gly Asn Glu Ser Phe LeuTrp Asp Cys His Ala Lys 1315 1320 1325 Pro Trp Gly Gln Ser Asp Cys GlyHis Lys Glu Asp Ala Gly Val Arg 1330 1335 1340 Cys Ser Gly Gln Ser LeuLys Ser Leu Asn Ala Ser Ser Gly His Leu 1345 1350 1355 1360 Ala Leu IleLeu Ser Ser Ile Phe Gly Leu Leu Leu Leu Val Leu Phe 1365 1370 1375 IleLeu Phe Leu Thr Trp Cys Arg Val Gln Lys Gln Lys His Leu Pro 1380 13851390 Leu Arg Val Ser Thr Arg Arg Arg Gly Ser Leu Glu Glu Asn Leu Phe1395 1400 1405 His Glu Met Glu Thr Cys Leu Lys Arg Glu Asp Pro His GlyThr Arg 1410 1415 1420 Thr Ser Asp Asp Thr Pro Asn His Gly Cys Glu AspAla Ser Asp Thr 1425 1430 1435 1440 Ser Leu Leu Gly Val Leu Pro Ala SerGlu Ala Thr Lys 1445 1450 12 40 PRT Homo sapiens 12 Met Met Leu Pro GlnAsn Ser Trp His Ile Asp Phe Gly Arg Cys Cys 1 5 10 15 Cys His Gln AsnLeu Phe Ser Ala Val Val Thr Cys Ile Leu Leu Leu 20 25 30 Asn Ser Cys PheLeu Ile Ser Ser 35 40 13 1413 PRT Homo sapiens 13 Phe Asn Gly Thr AspLeu Glu Leu Arg Leu Val Asn Gly Asp Gly Pro 1 5 10 15 Cys Ser Gly ThrVal Glu Val Lys Phe Gln Gly Gln Trp Gly Thr Val 20 25 30 Cys Asp Asp GlyTrp Asn Thr Thr Ala Ser Thr Val Val Cys Lys Gln 35 40 45 Leu Gly Cys ProPhe Ser Phe Ala Met Phe Arg Phe Gly Gln Ala Val 50 55 60 Thr Arg His GlyLys Ile Trp Leu Asp Asp Val Ser Cys Tyr Gly Asn 65 70 75 80 Glu Ser AlaLeu Trp Glu Cys Gln His Arg Glu Trp Gly Ser His Asn 85 90 95 Cys Tyr HisGly Glu Asp Val Gly Val Asn Cys Tyr Gly Glu Ala Asn 100 105 110 Leu GlyLeu Arg Leu Val Asp Gly Asn Asn Ser Cys Ser Gly Arg Val 115 120 125 GluVal Lys Phe Gln Glu Arg Trp Gly Thr Ile Cys Asp Asp Gly Trp 130 135 140Asn Leu Asn Thr Ala Ala Val Val Cys Arg Gln Leu Gly Cys Pro Ser 145 150155 160 Ser Phe Ile Ser Ser Gly Val Val Asn Ser Pro Ala Val Leu Arg Pro165 170 175 Ile Trp Leu Asp Asp Ile Leu Cys Gln Gly Asn Glu Leu Ala LeuTrp 180 185 190 Asn Cys Arg His Arg Gly Trp Gly Asn His Asp Cys Ser HisAsn Glu 195 200 205 Asp Val Thr Leu Thr Cys Tyr Asp Ser Ser Asp Leu GluLeu Arg Leu 210 215 220 Val Gly Gly Thr Asn Arg Cys Met Gly Arg Val GluLeu Lys Ile Gln 225 230 235 240 Gly Arg Trp Gly Thr Val Cys His His LysTrp Asn Asn Ala Ala Ala 245 250 255 Asp Val Val Cys Lys Gln Leu Gly CysGly Thr Ala Leu His Phe Ala 260 265 270 Gly Leu Pro His Leu Gln Ser GlySer Asp Val Val Trp Leu Asp Gly 275 280 285 Val Ser Cys Ser Gly Asn GluSer Phe Leu Trp Asp Cys Arg His Ser 290 295 300 Gly Thr Val Asn Phe AspCys Leu His Gln Asn Asp Val Ser Val Ile 305 310 315 320 Cys Ser Asp GlyAla Asp Leu Glu Leu Arg Leu Ala Asp Gly Ser Asn 325 330 335 Asn Cys SerGly Arg Val Glu Val Arg Ile His Glu Gln Trp Trp Thr 340 345 350 Ile CysAsp Gln Asn Trp Lys Asn Glu Gln Ala Leu Val Val Cys Lys 355 360 365 GlnLeu Gly Cys Pro Phe Ser Val Phe Gly Ser Arg Arg Ala Lys Pro 370 375 380Ser Asn Glu Ala Arg Asp Ile Trp Ile Asn Ser Ile Ser Cys Thr Gly 385 390395 400 Asn Glu Ser Ala Leu Trp Asp Cys Thr Tyr Asp Gly Lys Ala Lys Arg405 410 415 Thr Cys Phe Arg Arg Ser Asp Ala Gly Val Ile Cys Ser Asp LysAla 420 425 430 Asp Leu Asp Leu Arg Leu Val Gly Ala His Ser Pro Cys TyrGly Arg 435 440 445 Leu Glu Val Lys Tyr Gln Gly Glu Trp Gly Thr Val CysHis Asp Arg 450 455 460 Trp Ser Thr Arg Asn Ala Ala Val Val Cys Lys GlnLeu Gly Cys Gly 465 470 475 480 Lys Pro Met His Val Phe Gly Met Thr TyrPhe Lys Glu Ala Ser Gly 485 490 495 Pro Ile Trp Leu Asp Asp Val Ser CysIle Gly Asn Glu Ser Asn Ile 500 505 510 Trp Asp Cys Glu His Ser Gly TrpGly Lys His Asn Cys Val His Arg 515 520 525 Glu Asp Val Ile Val Thr CysSer Gly Asp Ala Thr Trp Gly Leu Arg 530 535 540 Leu Val Gly Gly Ser AsnArg Cys Ser Gly Arg Leu Glu Val Tyr Phe 545 550 555 560 Gln Gly Arg TrpGly Thr Val Cys Asp Asp Gly Trp Asn Ser Lys Ala 565 570 575 Ala Ala ValVal Cys Ser Gln Leu Asp Cys Pro Ser Ser Ile Ile Gly 580 585 590 Met GlyLeu Gly Asn Ala Ser Thr Gly Tyr Gly Lys Ile Trp Leu Asp 595 600 605 AspVal Ser Cys Asp Gly Asp Glu Ser Asp Leu Trp Ser Cys Arg Asn 610 615 620Ser Gly Trp Gly Asn Asn Asp Cys Ser His Ser Glu Asp Val Gly Val 625 630635 640 Ile Cys Ser Asp Ala Ser Asp Met Glu Leu Arg Leu Val Gly Gly Ser645 650 655 Ser Arg Cys Ala Gly Lys Val Glu Val Asn Val Gln Gly Ala ValGly 660 665 670 Ile Leu Cys Ala Asn Gly Trp Gly Met Asn Ile Ala Glu ValVal Cys 675 680 685 Arg Gln Leu Glu Cys Gly Ser Ala Ile Arg Val Ser ArgGlu Pro His 690 695 700 Phe Thr Glu Arg Thr Leu His Ile Leu Met Ser AsnSer Gly Cys Thr 705 710 715 720 Gly Gly Glu Ala Ser Leu Trp Asp Cys IleArg Trp Glu Trp Lys Gln 725 730 735 Thr Ala Cys His Leu Asn Met Glu AlaSer Leu Ile Cys Ser Ala His 740 745 750 Arg Gln Pro Arg Leu Val Gly AlaAsp Met Pro Cys Ser Gly Arg Val 755 760 765 Glu Val Lys His Ala Asp ThrTrp Arg Ser Val Cys Asp Ser Asp Phe 770 775 780 Ser Leu His Ala Ala AsnVal Leu Cys Arg Glu Leu Asn Cys Gly Asp 785 790 795 800 Ala Ile Ser LeuSer Val Gly Asp His Phe Gly Lys Gly Asn Gly Leu 805 810 815 Thr Trp AlaGlu Lys Phe Gln Cys Glu Gly Ser Glu Thr His Leu Ala 820 825 830 Leu CysPro Ile Val Gln His Pro Glu Asp Thr Cys Ile His Ser Arg 835 840 845 GluVal Gly Val Val Cys Ser Arg Tyr Thr Asp Val Arg Leu Val Asn 850 855 860Gly Lys Ser Gln Cys Asp Gly Gln Val Glu Ile Asn Val Leu Gly His 865 870875 880 Trp Gly Ser Leu Cys Asp Thr His Trp Asp Pro Glu Asp Ala Arg Val885 890 895 Leu Cys Arg Gln Leu Ser Cys Gly Thr Ala Leu Ser Thr Thr GlyGly 900 905 910 Lys Tyr Ile Gly Glu Arg Ser Val Arg Val Trp Gly His ArgPhe His 915 920 925 Cys Leu Gly Asn Glu Ser Leu Leu Asp Asn Cys Gln MetThr Val Leu 930 935 940 Gly Ala Pro Pro Cys Ile His Gly Asn Thr Val SerVal Ile Cys Thr 945 950 955 960 Gly Ser Leu Thr Gln Pro Leu Phe Pro CysLeu Ala Asn Val Ser Asp 965 970 975 Pro Tyr Leu Ser Ala Val Pro Glu GlySer Ala Leu Ile Cys Leu Glu 980 985 990 Asp Lys Arg Leu Arg Leu Val AspGly Asp Ser Arg Cys Ala Gly Arg 995 1000 1005 Val Glu Ile Tyr His AspGly Phe Trp Gly Thr Ile Cys Asp Asp Gly 1010 1015 1020 Trp Asp Leu SerAsp Ala His Val Val Cys Gln Lys Leu Gly Cys Gly 1025 1030 1035 1040 ValAla Phe Asn Ala Thr Val Ser Ala His Phe Gly Glu Gly Ser Gly 1045 10501055 Pro Ile Trp Leu Asp Asp Leu Asn Cys Thr Gly Thr Glu Ser His Leu1060 1065 1070 Trp Gln Cys Pro Ser Arg Gly Trp Gly Gln His Asp Cys ArgHis Lys 1075 1080 1085 Glu Asp Ala Gly Val Ile Cys Ser Glu Phe Thr AlaLeu Arg Leu Tyr 1090 1095 1100 Ser Glu Thr Glu Thr Glu Ser Cys Ala GlyArg Leu Glu Val Phe Tyr 1105 1110 1115 1120 Asn Gly Thr Trp Gly Ser ValGly Arg Arg Asn Ile Thr Thr Ala Ile 1125 1130 1135 Ala Gly Ile Val CysArg Gln Leu Gly Cys Gly Glu Asn Gly Val Val 1140 1145 1150 Ser Leu AlaPro Leu Ser Lys Thr Gly Ser Gly Phe Met Trp Val Asp 1155 1160 1165 AspIle Gln Cys Pro Lys Thr His Ile Ser Ile Trp Gln Cys Leu Ser 1170 11751180 Ala Pro Trp Glu Arg Arg Ile Ser Ser Pro Ala Glu Glu Thr Trp Ile1185 1190 1195 1200 Thr Cys Glu Asp Arg Ile Arg Val Arg Gly Gly Asp ThrGlu Cys Ser 1205 1210 1215 Gly Arg Val Glu Ile Trp His Ala Gly Ser TrpGly Thr Val Cys Asp 1220 1225 1230 Asp Ser Trp Asp Leu Ala Glu Ala GluVal Val Cys Gln Gln Leu Gly 1235 1240 1245 Cys Gly Ser Ala Leu Ala AlaLeu Arg Asp Ala Ser Phe Gly Gln Gly 1250 1255 1260 Thr Gly Thr Ile TrpLeu Asp Asp Met Arg Cys Lys Gly Asn Glu Ser 1265 1270 1275 1280 Phe LeuTrp Asp Cys His Ala Lys Pro Trp Gly Gln Ser Asp Cys Gly 1285 1290 1295His Lys Glu Asp Ala Gly Val Arg Cys Ser Gly Gln Ser Leu Lys Ser 13001305 1310 Leu Asn Ala Ser Ser Gly His Leu Ala Leu Ile Leu Ser Ser IlePhe 1315 1320 1325 Gly Leu Leu Leu Leu Val Leu Phe Ile Leu Phe Leu ThrTrp Cys Arg 1330 1335 1340 Val Gln Lys Gln Lys His Leu Pro Leu Arg ValSer Thr Arg Arg Arg 1345 1350 1355 1360 Gly Ser Leu Glu Glu Asn Leu PheHis Glu Met Glu Thr Cys Leu Lys 1365 1370 1375 Arg Glu Asp Pro His GlyThr Arg Thr Ser Asp Asp Thr Pro Asn His 1380 1385 1390 Gly Cys Glu AspAla Ser Asp Thr Ser Leu Leu Gly Val Leu Pro Ala 1395 1400 1405 Ser GluAla Thr Lys 1410 14 1319 PRT Homo sapiens 14 Phe Asn Gly Thr Asp Leu GluLeu Arg Leu Val Asn Gly Asp Gly Pro 1 5 10 15 Cys Ser Gly Thr Val GluVal Lys Phe Gln Gly Gln Trp Gly Thr Val 20 25 30 Cys Asp Asp Gly Trp AsnThr Thr Ala Ser Thr Val Val Cys Lys Gln 35 40 45 Leu Gly Cys Pro Phe SerPhe Ala Met Phe Arg Phe Gly Gln Ala Val 50 55 60 Thr Arg His Gly Lys IleTrp Leu Asp Asp Val Ser Cys Tyr Gly Asn 65 70 75 80 Glu Ser Ala Leu TrpGlu Cys Gln His Arg Glu Trp Gly Ser His Asn 85 90 95 Cys Tyr His Gly GluAsp Val Gly Val Asn Cys Tyr Gly Glu Ala Asn 100 105 110 Leu Gly Leu ArgLeu Val Asp Gly Asn Asn Ser Cys Ser Gly Arg Val 115 120 125 Glu Val LysPhe Gln Glu Arg Trp Gly Thr Ile Cys Asp Asp Gly Trp 130 135 140 Asn LeuAsn Thr Ala Ala Val Val Cys Arg Gln Leu Gly Cys Pro Ser 145 150 155 160Ser Phe Ile Ser Ser Gly Val Val Asn Ser Pro Ala Val Leu Arg Pro 165 170175 Ile Trp Leu Asp Asp Ile Leu Cys Gln Gly Asn Glu Leu Ala Leu Trp 180185 190 Asn Cys Arg His Arg Gly Trp Gly Asn His Asp Cys Ser His Asn Glu195 200 205 Asp Val Thr Leu Thr Cys Tyr Asp Ser Ser Asp Leu Glu Leu ArgLeu 210 215 220 Val Gly Gly Thr Asn Arg Cys Met Gly Arg Val Glu Leu LysIle Gln 225 230 235 240 Gly Arg Trp Gly Thr Val Cys His His Lys Trp AsnAsn Ala Ala Ala 245 250 255 Asp Val Val Cys Lys Gln Leu Gly Cys Gly ThrAla Leu His Phe Ala 260 265 270 Gly Leu Pro His Leu Gln Ser Gly Ser AspVal Val Trp Leu Asp Gly 275 280 285 Val Ser Cys Ser Gly Asn Glu Ser PheLeu Trp Asp Cys Arg His Ser 290 295 300 Gly Thr Val Asn Phe Asp Cys LeuHis Gln Asn Asp Val Ser Val Ile 305 310 315 320 Cys Ser Asp Gly Ala AspLeu Glu Leu Arg Leu Ala Asp Gly Ser Asn 325 330 335 Asn Cys Ser Gly ArgVal Glu Val Arg Ile His Glu Gln Trp Trp Thr 340 345 350 Ile Cys Asp GlnAsn Trp Lys Asn Glu Gln Ala Leu Val Val Cys Lys 355 360 365 Gln Leu GlyCys Pro Phe Ser Val Phe Gly Ser Arg Arg Ala Lys Pro 370 375 380 Ser AsnGlu Ala Arg Asp Ile Trp Ile Asn Ser Ile Ser Cys Thr Gly 385 390 395 400Asn Glu Ser Ala Leu Trp Asp Cys Thr Tyr Asp Gly Lys Ala Lys Arg 405 410415 Thr Cys Phe Arg Arg Ser Asp Ala Gly Val Ile Cys Ser Asp Lys Ala 420425 430 Asp Leu Asp Leu Arg Leu Val Gly Ala His Ser Pro Cys Tyr Gly Arg435 440 445 Leu Glu Val Lys Tyr Gln Gly Glu Trp Gly Thr Val Cys His AspArg 450 455 460 Trp Ser Thr Arg Asn Ala Ala Val Val Cys Lys Gln Leu GlyCys Gly 465 470 475 480 Lys Pro Met His Val Phe Gly Met Thr Tyr Phe LysGlu Ala Ser Gly 485 490 495 Pro Ile Trp Leu Asp Asp Val Ser Cys Ile GlyAsn Glu Ser Asn Ile 500 505 510 Trp Asp Cys Glu His Ser Gly Trp Gly LysHis Asn Cys Val His Arg 515 520 525 Glu Asp Val Ile Val Thr Cys Ser GlyAsp Ala Thr Trp Gly Leu Arg 530 535 540 Leu Val Gly Gly Ser Asn Arg CysSer Gly Arg Leu Glu Val Tyr Phe 545 550 555 560 Gln Gly Arg Trp Gly ThrVal Cys Asp Asp Gly Trp Asn Ser Lys Ala 565 570 575 Ala Ala Val Val CysSer Gln Leu Asp Cys Pro Ser Ser Ile Ile Gly 580 585 590 Met Gly Leu GlyAsn Ala Ser Thr Gly Tyr Gly Lys Ile Trp Leu Asp 595 600 605 Asp Val SerCys Asp Gly Asp Glu Ser Asp Leu Trp Ser Cys Arg Asn 610 615 620 Ser GlyTrp Gly Asn Asn Asp Cys Ser His Ser Glu Asp Val Gly Val 625 630 635 640Ile Cys Ser Asp Ala Ser Asp Met Glu Leu Arg Leu Val Gly Gly Ser 645 650655 Ser Arg Cys Ala Gly Lys Val Glu Val Asn Val Gln Gly Ala Val Gly 660665 670 Ile Leu Cys Ala Asn Gly Trp Gly Met Asn Ile Ala Glu Val Val Cys675 680 685 Arg Gln Leu Glu Cys Gly Ser Ala Ile Arg Val Ser Arg Glu ProHis 690 695 700 Phe Thr Glu Arg Thr Leu His Ile Leu Met Ser Asn Ser GlyCys Thr 705 710 715 720 Gly Gly Glu Ala Ser Leu Trp Asp Cys Ile Arg TrpGlu Trp Lys Gln 725 730 735 Thr Ala Cys His Leu Asn Met Glu Ala Ser LeuIle Cys Ser Ala His 740 745 750 Arg Gln Pro Arg Leu Val Gly Ala Asp MetPro Cys Ser Gly Arg Val 755 760 765 Glu Val Lys His Ala Asp Thr Trp ArgSer Val Cys Asp Ser Asp Phe 770 775 780 Ser Leu His Ala Ala Asn Val LeuCys Arg Glu Leu Asn Cys Gly Asp 785 790 795 800 Ala Ile Ser Leu Ser ValGly Asp His Phe Gly Lys Gly Asn Gly Leu 805 810 815 Thr Trp Ala Glu LysPhe Gln Cys Glu Gly Ser Glu Thr His Leu Ala 820 825 830 Leu Cys Pro IleVal Gln His Pro Glu Asp Thr Cys Ile His Ser Arg 835 840 845 Glu Val GlyVal Val Cys Ser Arg Tyr Thr Asp Val Arg Leu Val Asn 850 855 860 Gly LysSer Gln Cys Asp Gly Gln Val Glu Ile Asn Val Leu Gly His 865 870 875 880Trp Gly Ser Leu Cys Asp Thr His Trp Asp Pro Glu Asp Ala Arg Val 885 890895 Leu Cys Arg Gln Leu Ser Cys Gly Thr Ala Leu Ser Thr Thr Gly Gly 900905 910 Lys Tyr Ile Gly Glu Arg Ser Val Arg Val Trp Gly His Arg Phe His915 920 925 Cys Leu Gly Asn Glu Ser Leu Leu Asp Asn Cys Gln Met Thr ValLeu 930 935 940 Gly Ala Pro Pro Cys Ile His Gly Asn Thr Val Ser Val IleCys Thr 945 950 955 960 Gly Ser Leu Thr Gln Pro Leu Phe Pro Cys Leu AlaAsn Val Ser Asp 965 970 975 Pro Tyr Leu Ser Ala Val Pro Glu Gly Ser AlaLeu Ile Cys Leu Glu 980 985 990 Asp Lys Arg Leu Arg Leu Val Asp Gly AspSer Arg Cys Ala Gly Arg 995 1000 1005 Val Glu Ile Tyr His Asp Gly PheTrp Gly Thr Ile Cys Asp Asp Gly 1010 1015 1020 Trp Asp Leu Ser Asp AlaHis Val Val Cys Gln Lys Leu Gly Cys Gly 1025 1030 1035 1040 Val Ala PheAsn Ala Thr Val Ser Ala His Phe Gly Glu Gly Ser Gly 1045 1050 1055 ProIle Trp Leu Asp Asp Leu Asn Cys Thr Gly Thr Glu Ser His Leu 1060 10651070 Trp Gln Cys Pro Ser Arg Gly Trp Gly Gln His Asp Cys Arg His Lys1075 1080 1085 Glu Asp Ala Gly Val Ile Cys Ser Glu Phe Thr Ala Leu ArgLeu Tyr 1090 1095 1100 Ser Glu Thr Glu Thr Glu Ser Cys Ala Gly Arg LeuGlu Val Phe Tyr 1105 1110 1115 1120 Asn Gly Thr Trp Gly Ser Val Gly ArgArg Asn Ile Thr Thr Ala Ile 1125 1130 1135 Ala Gly Ile Val Cys Arg GlnLeu Gly Cys Gly Glu Asn Gly Val Val 1140 1145 1150 Ser Leu Ala Pro LeuSer Lys Thr Gly Ser Gly Phe Met Trp Val Asp 1155 1160 1165 Asp Ile GlnCys Pro Lys Thr His Ile Ser Ile Trp Gln Cys Leu Ser 1170 1175 1180 AlaPro Trp Glu Arg Arg Ile Ser Ser Pro Ala Glu Glu Thr Trp Ile 1185 11901195 1200 Thr Cys Glu Asp Arg Ile Arg Val Arg Gly Gly Asp Thr Glu CysSer 1205 1210 1215 Gly Arg Val Glu Ile Trp His Ala Gly Ser Trp Gly ThrVal Cys Asp 1220 1225 1230 Asp Ser Trp Asp Leu Ala Glu Ala Glu Val ValCys Gln Gln Leu Gly 1235 1240 1245 Cys Gly Ser Ala Leu Ala Ala Leu ArgAsp Ala Ser Phe Gly Gln Gly 1250 1255 1260 Thr Gly Thr Ile Trp Leu AspAsp Met Arg Cys Lys Gly Asn Glu Ser 1265 1270 1275 1280 Phe Leu Trp AspCys His Ala Lys Pro Trp Gly Gln Ser Asp Cys Gly 1285 1290 1295 His LysGlu Asp Ala Gly Val Arg Cys Ser Gly Gln Ser Leu Lys Ser 1300 1305 1310Leu Asn Ala Ser Ser Gly His 1315 15 24 PRT Homo sapiens 15 Leu Ala LeuIle Leu Ser Ser Ile Phe Gly Leu Leu Leu Leu Val Leu 1 5 10 15 Phe IleLeu Phe Leu Thr Trp Cys 20 16 70 PRT Homo sapiens 16 Arg Val Gln Lys GlnLys His Leu Pro Leu Arg Val Ser Thr Arg Arg 1 5 10 15 Arg Gly Ser LeuGlu Glu Asn Leu Phe His Glu Met Glu Thr Cys Leu 20 25 30 Lys Arg Glu AspPro His Gly Thr Arg Thr Ser Asp Asp Thr Pro Asn 35 40 45 His Gly Cys GluAsp Ala Ser Asp Thr Ser Leu Leu Gly Val Leu Pro 50 55 60 Ala Ser Glu AlaThr Lys 65 70 17 3104 DNA Homo sapiens 17 gtcgacccac gcgtccggtctgtggctgag catggccctc ccagccctgg gcctggaccc 60 ctggagcctc ctgggccttttcctcttcca actgcttcag ctgctgctgc cgacgacgac 120 cgcgggggga ggcgggcaggggcccatgcc cagggtcaga tactatgcag gggatgaacg 180 tagggcactt agcttcttccaccagaaggg cctccaggat tttgacactc tgctcctgag 240 tggtgatgga aatactctctacgtgggggc tcgagaagcc attctggcct tggatatcca 300 ggatccaggg gtccccaggctaaagaacat gataccgtgg ccagccagtg acagaaaaaa 360 gagtgaatgt gcctttaagaagaagagcaa tgagacacag tgtttcaact tcatccgtgt 420 cctggtttct tacaatgtcacccatctcta cacctgcggc accttcgcct tcagccctgc 480 ttgtaccttc attgaacttcaagattccta cctgttgccc atctcggagg acaaggtcat 540 ggagggaaaa ggccaaagcccctttgaccc cgctcacaag catacggctg tcttggtgga 600 tgggatgctc tattctggtactatgaacaa cttcctgggc agtgagccca tcctgatgcg 660 cacactggga tcccagcctgtcctcaagac cgacaacttc ctccgctggc tgcatcatga 720 cgcctccttt gtggcagccatcccttcgac ccaggtcgtc tacttcttct tcgaggagac 780 agccagcgag tttgacttctttgagaggct ccacacatcg cgggtggcta gagtctgcaa 840 gaatgacgtg ggcggcgaaaagctgctgca gaagaagtgg accaccttcc tgaaggccca 900 gctgctctgc acccagccggggcagctgcc cttcaacgtc atccgccacg cggtcctgct 960 ccccgccgat tctcccacagctccccacat ctacgcagtc ttcacctccc agtggcaggt 1020 tggcgggacc aggagctctgcggtttgtgc cttctctctc ttggacattg aacgtgtctt 1080 taaggggaaa tacaaagagttgaacaaaga aacttcacgc tggactactt ataggggccc 1140 tgagaccaac ccccggccaggcagttgctc agtgggcccc tcctctgata aggccctgac 1200 cttcatgaag gaccatttcctgatggatga gcaagtggtg gggacgcccc tgctggtgaa 1260 atctggcgtg gagtatacacggcttgcagt ggagacagcc cagggccttg atgggcacag 1320 ccatcttgtc atgtacctgggaaccaccac agggtcgctc cacaaggctg tggtaagtgg 1380 ggacagcagt gctcatctggtggaagagat tcagctgttc cctgaccctg aacctgttcg 1440 caacctgcag ctggcccccacccagggtgc agtgtttgta ggcttctcag gaggtgtctg 1500 gagggtgccc cgagccaactgtagtgtcta tgagagctgt gtggactgtg tccttgcccg 1560 ggacccccac tgtgcctgggaccctgagtc ccgaacctgt tgcctcctgt ctgcccccaa 1620 cctgaactcc tggaagcaggacatggagcg ggggaaccca gagtgggcat gtgccagtgg 1680 ccccatgagc aggagccttcggcctcagag ccgcccgcaa atcattaaag aagtcctggc 1740 tgtccccaac tccatcctggagctcccctg cccccacctg tcagccttgg cctcttatta 1800 ttggagtcat ggcccagcagcagtcccaga agcctcttcc actgtctaca atggctccct 1860 cttgctgata gtgcaggatggagttggggg tctctaccag tgctgggcaa ctgagaatgg 1920 cttttcatac cctgtgatctcctactgggt ggacagccag gaccagaccc tggccctgga 1980 tcctgaactg gcaggcatcccccgggagca tgtgaaggtc ccgttgacca gggtcagtgg 2040 tggggccgcc ctggctgcccagcagtccta ctggccccac tttgtcactg tcactgtcct 2100 ctttgcctta gtgctttcaggagccctcat catcctcgtg gcctccccat tgagagcact 2160 ccgggctcgg ggcaaggttcagggctgtga gaccctgcgc cctggggaga aggccccgtt 2220 aagcagagag caacacctccagtctcccaa ggaatgcagg acctctgcca gtgatgtgga 2280 cgctgacaac aactgcctaggcactgaggt agcttaaact ctaggcacag gccggggctg 2340 cggtgcaggc acctggccatgctggctggg cggcccaagc acagccctga ctaggatgac 2400 agcagcacaa aagaccacctttctcccctg agaggagctt ctgctactct gcatcactga 2460 tgacactcag cagggtgatgcacagcagtc tgcctcccct atgggactcc cttctaccaa 2520 gcacatgagc tctctaacagggtgggggct acccccagac ctgctcctac actgatattg 2580 aagaacctgg agaggatccttcagttctgg ccattccagg gaccctccag aaacacagtg 2640 tttcaagaga tcctaaaaaaacctgcctgt cccaggaccc tatggtaatg aacaccaaac 2700 atctaaacaa tcatatgctaacatgccact cctggaaact ccactctgaa gctgccgctt 2760 tggacaccaa cactcccttctcccagggtc atgcagggat ctgctccctc ctgcttccct 2820 taccagtcgt gcaccgctgactcccaggaa gtctttcctg aagtctgacc acctttcttc 2880 ttgcttcagt tggggcagactctgatccct tctgccctgg cagaatggca ggggtaatct 2940 gagccttctt cactcctttaccctagctga ccccttcacc tctccccctc ccttttcctt 3000 tgttttggga ttcagaaaactgcttgtcag agactgttta ttttttatta aaaatataag 3060 gcttaaaaaa aaaaaaaaaaaaaaaaaaaa aaaagggcgg ccgc 3104 18 2283 DNA Homo sapiens 18 atggccctcccagccctggg cctggacccc tggagcctcc tgggcctttt cctcttccaa 60 ctgcttcagctgctgctgcc gacgacgacc gcggggggag gcgggcaggg gcccatgccc 120 agggtcagatactatgcagg ggatgaacgt agggcactta gcttcttcca ccagaagggc 180 ctccaggattttgacactct gctcctgagt ggtgatggaa atactctcta cgtgggggct 240 cgagaagccattctggcctt ggatatccag gatccagggg tccccaggct aaagaacatg 300 ataccgtggccagccagtga cagaaaaaag agtgaatgtg cctttaagaa gaagagcaat 360 gagacacagtgtttcaactt catccgtgtc ctggtttctt acaatgtcac ccatctctac 420 acctgcggcaccttcgcctt cagccctgct tgtaccttca ttgaacttca agattcctac 480 ctgttgcccatctcggagga caaggtcatg gagggaaaag gccaaagccc ctttgacccc 540 gctcacaagcatacggctgt cttggtggat gggatgctct attctggtac tatgaacaac 600 ttcctgggcagtgagcccat cctgatgcgc acactgggat cccagcctgt cctcaagacc 660 gacaacttcctccgctggct gcatcatgac gcctcctttg tggcagccat cccttcgacc 720 caggtcgtctacttcttctt cgaggagaca gccagcgagt ttgacttctt tgagaggctc 780 cacacatcgcgggtggctag agtctgcaag aatgacgtgg gcggcgaaaa gctgctgcag 840 aagaagtggaccaccttcct gaaggcccag ctgctctgca cccagccggg gcagctgccc 900 ttcaacgtcatccgccacgc ggtcctgctc cccgccgatt ctcccacagc tccccacatc 960 tacgcagtcttcacctccca gtggcaggtt ggcgggacca ggagctctgc ggtttgtgcc 1020 ttctctctcttggacattga acgtgtcttt aaggggaaat acaaagagtt gaacaaagaa 1080 acttcacgctggactactta taggggccct gagaccaacc cccggccagg cagttgctca 1140 gtgggcccctcctctgataa ggccctgacc ttcatgaagg accatttcct gatggatgag 1200 caagtggtggggacgcccct gctggtgaaa tctggcgtgg agtatacacg gcttgcagtg 1260 gagacagcccagggccttga tgggcacagc catcttgtca tgtacctggg aaccaccaca 1320 gggtcgctccacaaggctgt ggtaagtggg gacagcagtg ctcatctggt ggaagagatt 1380 cagctgttccctgaccctga acctgttcgc aacctgcagc tggcccccac ccagggtgca 1440 gtgtttgtaggcttctcagg aggtgtctgg agggtgcccc gagccaactg tagtgtctat 1500 gagagctgtgtggactgtgt ccttgcccgg gacccccact gtgcctggga ccctgagtcc 1560 cgaacctgttgcctcctgtc tgcccccaac ctgaactcct ggaagcagga catggagcgg 1620 gggaacccagagtgggcatg tgccagtggc cccatgagca ggagccttcg gcctcagagc 1680 cgcccgcaaatcattaaaga agtcctggct gtccccaact ccatcctgga gctcccctgc 1740 ccccacctgtcagccttggc ctcttattat tggagtcatg gcccagcagc agtcccagaa 1800 gcctcttccactgtctacaa tggctccctc ttgctgatag tgcaggatgg agttgggggt 1860 ctctaccagtgctgggcaac tgagaatggc ttttcatacc ctgtgatctc ctactgggtg 1920 gacagccaggaccagaccct ggccctggat cctgaactgg caggcatccc ccgggagcat 1980 gtgaaggtcccgttgaccag ggtcagtggt ggggccgccc tggctgccca gcagtcctac 2040 tggccccactttgtcactgt cactgtcctc tttgccttag tgctttcagg agccctcatc 2100 atcctcgtggcctccccatt gagagcactc cgggctcggg gcaaggttca gggctgtgag 2160 accctgcgccctggggagaa ggccccgtta agcagagagc aacacctcca gtctcccaag 2220 gaatgcaggacctctgccag tgatgtggac gctgacaaca actgcctagg cactgaggta 2280 gct 2283 19761 PRT Homo sapiens 19 Met Ala Leu Pro Ala Leu Gly Leu Asp Pro Trp SerLeu Leu Gly Leu 1 5 10 15 Phe Leu Phe Gln Leu Leu Gln Leu Leu Leu ProThr Thr Thr Ala Gly 20 25 30 Gly Gly Gly Gln Gly Pro Met Pro Arg Val ArgTyr Tyr Ala Gly Asp 35 40 45 Glu Arg Arg Ala Leu Ser Phe Phe His Gln LysGly Leu Gln Asp Phe 50 55 60 Asp Thr Leu Leu Leu Ser Gly Asp Gly Asn ThrLeu Tyr Val Gly Ala 65 70 75 80 Arg Glu Ala Ile Leu Ala Leu Asp Ile GlnAsp Pro Gly Val Pro Arg 85 90 95 Leu Lys Asn Met Ile Pro Trp Pro Ala SerAsp Arg Lys Lys Ser Glu 100 105 110 Cys Ala Phe Lys Lys Lys Ser Asn GluThr Gln Cys Phe Asn Phe Ile 115 120 125 Arg Val Leu Val Ser Tyr Asn ValThr His Leu Tyr Thr Cys Gly Thr 130 135 140 Phe Ala Phe Ser Pro Ala CysThr Phe Ile Glu Leu Gln Asp Ser Tyr 145 150 155 160 Leu Leu Pro Ile SerGlu Asp Lys Val Met Glu Gly Lys Gly Gln Ser 165 170 175 Pro Phe Asp ProAla His Lys His Thr Ala Val Leu Val Asp Gly Met 180 185 190 Leu Tyr SerGly Thr Met Asn Asn Phe Leu Gly Ser Glu Pro Ile Leu 195 200 205 Met ArgThr Leu Gly Ser Gln Pro Val Leu Lys Thr Asp Asn Phe Leu 210 215 220 ArgTrp Leu His His Asp Ala Ser Phe Val Ala Ala Ile Pro Ser Thr 225 230 235240 Gln Val Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp Phe 245250 255 Phe Glu Arg Leu His Thr Ser Arg Val Ala Arg Val Cys Lys Asn Asp260 265 270 Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe LeuLys 275 280 285 Ala Gln Leu Leu Cys Thr Gln Pro Gly Gln Leu Pro Phe AsnVal Ile 290 295 300 Arg His Ala Val Leu Leu Pro Ala Asp Ser Pro Thr AlaPro His Ile 305 310 315 320 Tyr Ala Val Phe Thr Ser Gln Trp Gln Val GlyGly Thr Arg Ser Ser 325 330 335 Ala Val Cys Ala Phe Ser Leu Leu Asp IleGlu Arg Val Phe Lys Gly 340 345 350 Lys Tyr Lys Glu Leu Asn Lys Glu ThrSer Arg Trp Thr Thr Tyr Arg 355 360 365 Gly Pro Glu Thr Asn Pro Arg ProGly Ser Cys Ser Val Gly Pro Ser 370 375 380 Ser Asp Lys Ala Leu Thr PheMet Lys Asp His Phe Leu Met Asp Glu 385 390 395 400 Gln Val Val Gly ThrPro Leu Leu Val Lys Ser Gly Val Glu Tyr Thr 405 410 415 Arg Leu Ala ValGlu Thr Ala Gln Gly Leu Asp Gly His Ser His Leu 420 425 430 Val Met TyrLeu Gly Thr Thr Thr Gly Ser Leu His Lys Ala Val Val 435 440 445 Ser GlyAsp Ser Ser Ala His Leu Val Glu Glu Ile Gln Leu Phe Pro 450 455 460 AspPro Glu Pro Val Arg Asn Leu Gln Leu Ala Pro Thr Gln Gly Ala 465 470 475480 Val Phe Val Gly Phe Ser Gly Gly Val Trp Arg Val Pro Arg Ala Asn 485490 495 Cys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala Arg Asp Pro500 505 510 His Cys Ala Trp Asp Pro Glu Ser Arg Thr Cys Cys Leu Leu SerAla 515 520 525 Pro Asn Leu Asn Ser Trp Lys Gln Asp Met Glu Arg Gly AsnPro Glu 530 535 540 Trp Ala Cys Ala Ser Gly Pro Met Ser Arg Ser Leu ArgPro Gln Ser 545 550 555 560 Arg Pro Gln Ile Ile Lys Glu Val Leu Ala ValPro Asn Ser Ile Leu 565 570 575 Glu Leu Pro Cys Pro His Leu Ser Ala LeuAla Ser Tyr Tyr Trp Ser 580 585 590 His Gly Pro Ala Ala Val Pro Glu AlaSer Ser Thr Val Tyr Asn Gly 595 600 605 Ser Leu Leu Leu Ile Val Gln AspGly Val Gly Gly Leu Tyr Gln Cys 610 615 620 Trp Ala Thr Glu Asn Gly PheSer Tyr Pro Val Ile Ser Tyr Trp Val 625 630 635 640 Asp Ser Gln Asp GlnThr Leu Ala Leu Asp Pro Glu Leu Ala Gly Ile 645 650 655 Pro Arg Glu HisVal Lys Val Pro Leu Thr Arg Val Ser Gly Gly Ala 660 665 670 Ala Leu AlaAla Gln Gln Ser Tyr Trp Pro His Phe Val Thr Val Thr 675 680 685 Val LeuPhe Ala Leu Val Leu Ser Gly Ala Leu Ile Ile Leu Val Ala 690 695 700 SerPro Leu Arg Ala Leu Arg Ala Arg Gly Lys Val Gln Gly Cys Glu 705 710 715720 Thr Leu Arg Pro Gly Glu Lys Ala Pro Leu Ser Arg Glu Gln His Leu 725730 735 Gln Ser Pro Lys Glu Cys Arg Thr Ser Ala Ser Asp Val Asp Ala Asp740 745 750 Asn Asn Cys Leu Gly Thr Glu Val Ala 755 760 20 31 PRT Homosapiens 20 Met Ala Leu Pro Ala Leu Gly Leu Asp Pro Trp Ser Leu Leu GlyLeu 1 5 10 15 Phe Leu Phe Gln Leu Leu Gln Leu Leu Leu Pro Thr Thr ThrAla 20 25 30 21 730 PRT Homo sapiens 21 Gly Gly Gly Gly Gln Gly Pro MetPro Arg Val Arg Tyr Tyr Ala Gly 1 5 10 15 Asp Glu Arg Arg Ala Leu SerPhe Phe His Gln Lys Gly Leu Gln Asp 20 25 30 Phe Asp Thr Leu Leu Leu SerGly Asp Gly Asn Thr Leu Tyr Val Gly 35 40 45 Ala Arg Glu Ala Ile Leu AlaLeu Asp Ile Gln Asp Pro Gly Val Pro 50 55 60 Arg Leu Lys Asn Met Ile ProTrp Pro Ala Ser Asp Arg Lys Lys Ser 65 70 75 80 Glu Cys Ala Phe Lys LysLys Ser Asn Glu Thr Gln Cys Phe Asn Phe 85 90 95 Ile Arg Val Leu Val SerTyr Asn Val Thr His Leu Tyr Thr Cys Gly 100 105 110 Thr Phe Ala Phe SerPro Ala Cys Thr Phe Ile Glu Leu Gln Asp Ser 115 120 125 Tyr Leu Leu ProIle Ser Glu Asp Lys Val Met Glu Gly Lys Gly Gln 130 135 140 Ser Pro PheAsp Pro Ala His Lys His Thr Ala Val Leu Val Asp Gly 145 150 155 160 MetLeu Tyr Ser Gly Thr Met Asn Asn Phe Leu Gly Ser Glu Pro Ile 165 170 175Leu Met Arg Thr Leu Gly Ser Gln Pro Val Leu Lys Thr Asp Asn Phe 180 185190 Leu Arg Trp Leu His His Asp Ala Ser Phe Val Ala Ala Ile Pro Ser 195200 205 Thr Gln Val Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp210 215 220 Phe Phe Glu Arg Leu His Thr Ser Arg Val Ala Arg Val Cys LysAsn 225 230 235 240 Asp Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp ThrThr Phe Leu 245 250 255 Lys Ala Gln Leu Leu Cys Thr Gln Pro Gly Gln LeuPro Phe Asn Val 260 265 270 Ile Arg His Ala Val Leu Leu Pro Ala Asp SerPro Thr Ala Pro His 275 280 285 Ile Tyr Ala Val Phe Thr Ser Gln Trp GlnVal Gly Gly Thr Arg Ser 290 295 300 Ser Ala Val Cys Ala Phe Ser Leu LeuAsp Ile Glu Arg Val Phe Lys 305 310 315 320 Gly Lys Tyr Lys Glu Leu AsnLys Glu Thr Ser Arg Trp Thr Thr Tyr 325 330 335 Arg Gly Pro Glu Thr AsnPro Arg Pro Gly Ser Cys Ser Val Gly Pro 340 345 350 Ser Ser Asp Lys AlaLeu Thr Phe Met Lys Asp His Phe Leu Met Asp 355 360 365 Glu Gln Val ValGly Thr Pro Leu Leu Val Lys Ser Gly Val Glu Tyr 370 375 380 Thr Arg LeuAla Val Glu Thr Ala Gln Gly Leu Asp Gly His Ser His 385 390 395 400 LeuVal Met Tyr Leu Gly Thr Thr Thr Gly Ser Leu His Lys Ala Val 405 410 415Val Ser Gly Asp Ser Ser Ala His Leu Val Glu Glu Ile Gln Leu Phe 420 425430 Pro Asp Pro Glu Pro Val Arg Asn Leu Gln Leu Ala Pro Thr Gln Gly 435440 445 Ala Val Phe Val Gly Phe Ser Gly Gly Val Trp Arg Val Pro Arg Ala450 455 460 Asn Cys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala ArgAsp 465 470 475 480 Pro His Cys Ala Trp Asp Pro Glu Ser Arg Thr Cys CysLeu Leu Ser 485 490 495 Ala Pro Asn Leu Asn Ser Trp Lys Gln Asp Met GluArg Gly Asn Pro 500 505 510 Glu Trp Ala Cys Ala Ser Gly Pro Met Ser ArgSer Leu Arg Pro Gln 515 520 525 Ser Arg Pro Gln Ile Ile Lys Glu Val LeuAla Val Pro Asn Ser Ile 530 535 540 Leu Glu Leu Pro Cys Pro His Leu SerAla Leu Ala Ser Tyr Tyr Trp 545 550 555 560 Ser His Gly Pro Ala Ala ValPro Glu Ala Ser Ser Thr Val Tyr Asn 565 570 575 Gly Ser Leu Leu Leu IleVal Gln Asp Gly Val Gly Gly Leu Tyr Gln 580 585 590 Cys Trp Ala Thr GluAsn Gly Phe Ser Tyr Pro Val Ile Ser Tyr Trp 595 600 605 Val Asp Ser GlnAsp Gln Thr Leu Ala Leu Asp Pro Glu Leu Ala Gly 610 615 620 Ile Pro ArgGlu His Val Lys Val Pro Leu Thr Arg Val Ser Gly Gly 625 630 635 640 AlaAla Leu Ala Ala Gln Gln Ser Tyr Trp Pro His Phe Val Thr Val 645 650 655Thr Val Leu Phe Ala Leu Val Leu Ser Gly Ala Leu Ile Ile Leu Val 660 665670 Ala Ser Pro Leu Arg Ala Leu Arg Ala Arg Gly Lys Val Gln Gly Cys 675680 685 Glu Thr Leu Arg Pro Gly Glu Lys Ala Pro Leu Ser Arg Glu Gln His690 695 700 Leu Gln Ser Pro Lys Glu Cys Arg Thr Ser Ala Ser Asp Val AspAla 705 710 715 720 Asp Asn Asn Cys Leu Gly Thr Glu Val Ala 725 730 22652 PRT Homo sapiens 22 Gly Gly Gly Gly Gln Gly Pro Met Pro Arg Val ArgTyr Tyr Ala Gly 1 5 10 15 Asp Glu Arg Arg Ala Leu Ser Phe Phe His GlnLys Gly Leu Gln Asp 20 25 30 Phe Asp Thr Leu Leu Leu Ser Gly Asp Gly AsnThr Leu Tyr Val Gly 35 40 45 Ala Arg Glu Ala Ile Leu Ala Leu Asp Ile GlnAsp Pro Gly Val Pro 50 55 60 Arg Leu Lys Asn Met Ile Pro Trp Pro Ala SerAsp Arg Lys Lys Ser 65 70 75 80 Glu Cys Ala Phe Lys Lys Lys Ser Asn GluThr Gln Cys Phe Asn Phe 85 90 95 Ile Arg Val Leu Val Ser Tyr Asn Val ThrHis Leu Tyr Thr Cys Gly 100 105 110 Thr Phe Ala Phe Ser Pro Ala Cys ThrPhe Ile Glu Leu Gln Asp Ser 115 120 125 Tyr Leu Leu Pro Ile Ser Glu AspLys Val Met Glu Gly Lys Gly Gln 130 135 140 Ser Pro Phe Asp Pro Ala HisLys His Thr Ala Val Leu Val Asp Gly 145 150 155 160 Met Leu Tyr Ser GlyThr Met Asn Asn Phe Leu Gly Ser Glu Pro Ile 165 170 175 Leu Met Arg ThrLeu Gly Ser Gln Pro Val Leu Lys Thr Asp Asn Phe 180 185 190 Leu Arg TrpLeu His His Asp Ala Ser Phe Val Ala Ala Ile Pro Ser 195 200 205 Thr GlnVal Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp 210 215 220 PhePhe Glu Arg Leu His Thr Ser Arg Val Ala Arg Val Cys Lys Asn 225 230 235240 Asp Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe Leu 245250 255 Lys Ala Gln Leu Leu Cys Thr Gln Pro Gly Gln Leu Pro Phe Asn Val260 265 270 Ile Arg His Ala Val Leu Leu Pro Ala Asp Ser Pro Thr Ala ProHis 275 280 285 Ile Tyr Ala Val Phe Thr Ser Gln Trp Gln Val Gly Gly ThrArg Ser 290 295 300 Ser Ala Val Cys Ala Phe Ser Leu Leu Asp Ile Glu ArgVal Phe Lys 305 310 315 320 Gly Lys Tyr Lys Glu Leu Asn Lys Glu Thr SerArg Trp Thr Thr Tyr 325 330 335 Arg Gly Pro Glu Thr Asn Pro Arg Pro GlySer Cys Ser Val Gly Pro 340 345 350 Ser Ser Asp Lys Ala Leu Thr Phe MetLys Asp His Phe Leu Met Asp 355 360 365 Glu Gln Val Val Gly Thr Pro LeuLeu Val Lys Ser Gly Val Glu Tyr 370 375 380 Thr Arg Leu Ala Val Glu ThrAla Gln Gly Leu Asp Gly His Ser His 385 390 395 400 Leu Val Met Tyr LeuGly Thr Thr Thr Gly Ser Leu His Lys Ala Val 405 410 415 Val Ser Gly AspSer Ser Ala His Leu Val Glu Glu Ile Gln Leu Phe 420 425 430 Pro Asp ProGlu Pro Val Arg Asn Leu Gln Leu Ala Pro Thr Gln Gly 435 440 445 Ala ValPhe Val Gly Phe Ser Gly Gly Val Trp Arg Val Pro Arg Ala 450 455 460 AsnCys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala Arg Asp 465 470 475480 Pro His Cys Ala Trp Asp Pro Glu Ser Arg Thr Cys Cys Leu Leu Ser 485490 495 Ala Pro Asn Leu Asn Ser Trp Lys Gln Asp Met Glu Arg Gly Asn Pro500 505 510 Glu Trp Ala Cys Ala Ser Gly Pro Met Ser Arg Ser Leu Arg ProGln 515 520 525 Ser Arg Pro Gln Ile Ile Lys Glu Val Leu Ala Val Pro AsnSer Ile 530 535 540 Leu Glu Leu Pro Cys Pro His Leu Ser Ala Leu Ala SerTyr Tyr Trp 545 550 555 560 Ser His Gly Pro Ala Ala Val Pro Glu Ala SerSer Thr Val Tyr Asn 565 570 575 Gly Ser Leu Leu Leu Ile Val Gln Asp GlyVal Gly Gly Leu Tyr Gln 580 585 590 Cys Trp Ala Thr Glu Asn Gly Phe SerTyr Pro Val Ile Ser Tyr Trp 595 600 605 Val Asp Ser Gln Asp Gln Thr LeuAla Leu Asp Pro Glu Leu Ala Gly 610 615 620 Ile Pro Arg Glu His Val LysVal Pro Leu Thr Arg Val Ser Gly Gly 625 630 635 640 Ala Ala Leu Ala AlaGln Gln Ser Tyr Trp Pro His 645 650 23 21 PRT Homo sapiens 23 Phe ValThr Val Thr Val Leu Phe Ala Leu Val Leu Ser Gly Ala Leu 1 5 10 15 IleIle Leu Val Ala 20 24 57 PRT Homo sapiens 24 Ser Pro Leu Arg Ala Leu ArgAla Arg Gly Lys Val Gln Gly Cys Glu 1 5 10 15 Thr Leu Arg Pro Gly GluLys Ala Pro Leu Ser Arg Glu Gln His Leu 20 25 30 Gln Ser Pro Lys Glu CysArg Thr Ser Ala Ser Asp Val Asp Ala Asp 35 40 45 Asn Asn Cys Leu Gly ThrGlu Val Ala 50 55 25 2964 DNA Homo sapiens 25 gtcgacccac gcgtccgcggacgcgtgggg acggctcccg gctgcagtct gcccgcccgc 60 cccgcgcggg ggccgagtcgcgaagcgcgc ctgcgacccg gcgtccgggc gcgctggaga 120 ggacgcgagg agccatgaggcgccagcctg cgaaggtggc ggcgctgctg ctcgggctgc 180 tcttggagtg cacagaagccaaaaagcatt gctggtattt cgaaggactc tatccaacct 240 attatatatg ccgctcctacgaggactgct gtggctccag gtgctgtgtg cgggccctct 300 ccatacagag gctgtggtacttctggttcc ttctgatgat gggcgtgctt ttctgctgcg 360 gagccggctt cttcatccggaggcgcatgt accccccgcc gctgatcgag gagccagcct 420 tcaatgtgtc ctacaccaggcagcccccaa atcccggccc aggagcccag cagccggggc 480 cgccctatta cactgacccaggaggaccgg ggatgaaccc tgtcgggaat tccatggcaa 540 tggctttcca ggtcccacccaactcacccc aggggagtgt ggcctgcccg ccccctccag 600 cctactgcaa cacgcctccgcccccgtacg aacaggtagt gaaggccaag tagtggggtg 660 cccacgtgca agaggagagacaggagaggg cctttccctg gcctttctgt cttcgttgat 720 gttcacttcc aggaacggtctcgtgggctg ctaagggcag ttcctctgat atcctcacag 780 caagcacagc tctctttcaggctttccatg gagtacaata tatgaactca cactttgtct 840 cctctgttgc ttctgtttctgacgcagtct gtgctctcac atggtagtgt ggtgacagtc 900 cccgagggct gacgtccttacggtggcgtg accagatcta caggagagag actgagagga 960 agaaggcagt gctggaggtgcaggtggcat gtagaggggc caggccgagc atcccaggca 1020 agcatccttc tgcccgggtattaataggaa gccccatgcc gggcggctca gccgatgaag 1080 cagcagccga ctgagctgagcccagcaggt catctgctcc agcctgtcct ctcgtcagcc 1140 ttcctcttcc agaagctgttggagagacat tcaggagaga gcaagcccct tgtcatgttt 1200 ctgtctctgt tcatatcctaaagatagact tctcctgcac cgccagggaa gggtagcacg 1260 tgcagctctc accgcaggatggggcctaga atcaggcttg ccttggaggc ctgacagtga 1320 tctgacatcc actaagcaaatttatttaaa ttcatgggaa atcacttcct gccccaaact 1380 gagacattgc attttgtgagctcttggtct gatttggaga aaggactgtt acccattttt 1440 ttggtgtgtt tatggaagtgcatgtagagc gtcctgccct ttgaaatcag actgggtgtg 1500 tgtcttccct ggacatcactgcctctccag ggcattctca ggcccggggg tctccttccc 1560 tcaggcagct ccagtggtgggttctgaagg gtgctttcaa aacggggcac atctggctgg 1620 gaagtcacat ggactcttccagggagagag accagctgag gcgtctctct ctgaggttgt 1680 gttgggtcta agcgggtgtgtgctgggctc caaggaggag gagcttgctg ggaaaagaca 1740 ggagaagtac tgactcaactgcactgacca tgttgtcata attagaataa agaagaagtg 1800 gtcggaaatg cacattcctggataggaatc acagctcacc ccaggatctc acaggtagtc 1860 tcctgagtag ttgacggctagcggggagct agttccgccg catagttata gtgttgatgt 1920 gtgaacgctg acctgtcctgtgtgctaaga gctatgcagc ttagctgagg cgcctagatt 1980 actagatgtg ctgtatcacggggaatgagg tgggggtgct tattttttaa tgaactaatc 2040 agagcctctt gagaaattgttactcattga actggagcat caagacatct catggaagtg 2100 gatacggagt gatttggtgtccatgctttt cactctgagg acatttaatc ggagaacctc 2160 ctggggaatt ttgtgggagacacttgggaa caaaacagac accctgggaa tgcagttgca 2220 agcacagatg ctgccaccagtgtctctgac caccctggtg tgactgctga ctgccagcgt 2280 ggtacctccc atgctgcaggcctccatcta aatgagacaa caaagcacaa tgttcactgt 2340 ttacaaccaa gacaactgcgtgggtccaaa cactcctctt cctccaggtc atttgttttg 2400 catttttaat gtctttattttttgtaatga aaaagcacac taagctgccc ctggaatcgg 2460 gtgcagctga ataggcacccaaaagtccgt gactaaattt cgtttgtctt tttgatagca 2520 aattatgtta agagacagtgatggctaggg ctcaacaatt ttgtattccc atgtttgtgt 2580 gagacagagt ttgttttcccttgaacttgg ttagaattgt gctactgtga acgctgatcc 2640 tgcatatgga agtcccactttggtgacatt tcctggccat tcttgtttcc attgtgtgga 2700 tggtgggttg tgcccacttcctggagtgag acagctcctg gtgtgtagaa ttcccggagc 2760 gtccgtggtt cagagtaaacttgaagcaga tctgtgcatg cttttcctct gcaacaattg 2820 gctcgtttct cttttttgttctcttttgat aggatcctgt ttcctatgtg tgcaaaataa 2880 aaataaattt gggcaaaaaaaaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2940 aaaaaaaaaa aaaagggcggccgc 2964 26 516 DNA Homo sapiens 26 atgaggcgcc agcctgcgaa ggtggcggcgctgctgctcg ggctgctctt ggagtgcaca 60 gaagccaaaa agcattgctg gtatttcgaaggactctatc caacctatta tatatgccgc 120 tcctacgagg actgctgtgg ctccaggtgctgtgtgcggg ccctctccat acagaggctg 180 tggtacttct ggttccttct gatgatgggcgtgcttttct gctgcggagc cggcttcttc 240 atccggaggc gcatgtaccc cccgccgctgatcgaggagc cagccttcaa tgtgtcctac 300 accaggcagc ccccaaatcc cggcccaggagcccagcagc cggggccgcc ctattacact 360 gacccaggag gaccggggat gaaccctgtcgggaattcca tggcaatggc tttccaggtc 420 ccacccaact caccccaggg gagtgtggcctgcccgcccc ctccagccta ctgcaacacg 480 cctccgcccc cgtacgaaca ggtagtgaaggccaag 516 27 172 PRT Homo sapiens 27 Met Arg Arg Gln Pro Ala Lys ValAla Ala Leu Leu Leu Gly Leu Leu 1 5 10 15 Leu Glu Cys Thr Glu Ala LysLys His Cys Trp Tyr Phe Glu Gly Leu 20 25 30 Tyr Pro Thr Tyr Tyr Ile CysArg Ser Tyr Glu Asp Cys Cys Gly Ser 35 40 45 Arg Cys Cys Val Arg Ala LeuSer Ile Gln Arg Leu Trp Tyr Phe Trp 50 55 60 Phe Leu Leu Met Met Gly ValLeu Phe Cys Cys Gly Ala Gly Phe Phe 65 70 75 80 Ile Arg Arg Arg Met TyrPro Pro Pro Leu Ile Glu Glu Pro Ala Phe 85 90 95 Asn Val Ser Tyr Thr ArgGln Pro Pro Asn Pro Gly Pro Gly Ala Gln 100 105 110 Gln Pro Gly Pro ProTyr Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn 115 120 125 Pro Val Gly AsnSer Met Ala Met Ala Phe Gln Val Pro Pro Asn Ser 130 135 140 Pro Gln GlySer Val Ala Cys Pro Pro Pro Pro Ala Tyr Cys Asn Thr 145 150 155 160 ProPro Pro Pro Tyr Glu Gln Val Val Lys Ala Lys 165 170 28 22 PRT Homosapiens 28 Met Arg Arg Gln Pro Ala Lys Val Ala Ala Leu Leu Leu Gly LeuLeu 1 5 10 15 Leu Glu Cys Thr Glu Ala 20 29 150 PRT Homo sapiens 29 LysLys His Cys Trp Tyr Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1 5 10 15Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg Ala 20 25 30Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp Phe Leu Leu Met Met Gly 35 40 45Val Leu Phe Cys Cys Gly Ala Gly Phe Phe Ile Arg Arg Arg Met Tyr 50 55 60Pro Pro Pro Leu Ile Glu Glu Pro Ala Phe Asn Val Ser Tyr Thr Arg 65 70 7580 Gln Pro Pro Asn Pro Gly Pro Gly Ala Gln Gln Pro Gly Pro Pro Tyr 85 9095 Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn Pro Val Gly Asn Ser Met 100105 110 Ala Met Ala Phe Gln Val Pro Pro Asn Ser Pro Gln Gly Ser Val Ala115 120 125 Cys Pro Pro Pro Pro Ala Tyr Cys Asn Thr Pro Pro Pro Pro TyrGlu 130 135 140 Gln Val Val Lys Ala Lys 145 150 30 38 PRT Homo sapiens30 Lys Lys His Cys Trp Tyr Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 1 510 15 Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg Ala 2025 30 Leu Ser Ile Gln Arg Leu 35 31 21 PRT Homo sapiens 31 Trp Tyr PheTrp Phe Leu Leu Met Met Gly Val Leu Phe Cys Cys Gly 1 5 10 15 Ala GlyPhe Phe Ile 20 32 91 PRT Homo sapiens 32 Arg Arg Arg Met Tyr Pro Pro ProLeu Ile Glu Glu Pro Ala Phe Asn 1 5 10 15 Val Ser Tyr Thr Arg Gln ProPro Asn Pro Gly Pro Gly Ala Gln Gln 20 25 30 Pro Gly Pro Pro Tyr Tyr ThrAsp Pro Gly Gly Pro Gly Met Asn Pro 35 40 45 Val Gly Asn Ser Met Ala MetAla Phe Gln Val Pro Pro Asn Ser Pro 50 55 60 Gln Gly Ser Val Ala Cys ProPro Pro Pro Ala Tyr Cys Asn Thr Pro 65 70 75 80 Pro Pro Pro Tyr Glu GlnVal Val Lys Ala Lys 85 90 33 1980 DNA Homo sapiens 33 gtcgacccacgcgtccgcag ctttggacac ttcctctgct tgaggacacc ttgactaacc 60 tccaagggcaactaaaggat caagaaaggc ccagcacagc agaagatcag ctggatctag 120 ctcctgcaggagatgtgtac aaagacaatc ccagtcctct ggggatgttt cctcctgtgg 180 aatctctatgtctcatcctc tcagaccatt taccctggaa tcaaggcaag gattactcag 240 agggcacttgactatggtgt tcaagctgga atgaagatga ttgagcaaat gctaaaagaa 300 aagaaactcccagatttaag cggttctgag tctcttgaat ttctaaaagt tgattatgta 360 aactacaatttttcaaatat aaaaatcagt gccttttcat ttccaaatac ctcattggct 420 tttgtgcctggagtgggaat caaagcgcta accaaccatg gcactgccaa catcagcaca 480 gactgggggttcgagtctcc actttttgtt ctgtataact cctttgctga gcccatggag 540 aaacccattttaaagaactt aaatgaaatg ctctgtccca ttattgcaag tgaagtcaaa 600 gcgctaaatgccaacctcag cacactggag gttttaacca agattgacaa ctacactctg 660 ctggattactccctaatcag ttctccagaa attactgaga actaccttga cctgaacttg 720 aagggtgtattctacccact ggaaaacctc accgaccccc ccttctcacc agttcctttt 780 gtgctcccagaacgcagcaa ctccatgctc tacattggaa tcgccgagta tttctttaaa 840 tctgcgtcctttgctcattt cacagctggg gttttcaatc tcactctctc caccgaagag 900 atttccaaccattttgttca aaactctcaa ggccttggca acgtgctctc ccggattgca 960 gagatctacatcttgtccca gcccttcatg gtgaggatca tggccacaga gcctcccata 1020 atcaatctacaaccaggcaa tttcaccctg gacatccctg cctccatcat gatgctcacc 1080 caacccaagaactccacagt tgaaaccatc gtttccatgg acttcgttgc tagtaccagt 1140 gttggcctggttattttggg acaaagactg gtctgctcct tgtctctgaa cagattccgc 1200 cttgctttgccagagtccaa tcgcagcaac attgaggtct tgaggtttga aaatattcta 1260 tcgtccattcttcactttgg agtcctccca ctggccaatg caaaattgca gcaaggattt 1320 cctctgcccaatccacacaa attcttattc gtcaattcag atattgaagt tcttgagggt 1380 ttccttttgatttccaccga cctgaagtat gaaacatcct caaagcagca gccaagtttc 1440 cacgtatgggaaggtctgaa cctgataagc agacagtgga gggggaagtc agccccttga 1500 ttgccggtttgcaattcacc ccaggaagta aatggtcctt aatcctacaa ctactgtaaa 1560 cccagaagggaaagacagta cacactggaa ttgtaaagcc cttgtgaatt gcttaggcag 1620 aaagttttctttcttaagcc ttcaggaacc cagaataagg cagactctgt taaagggata 1680 aatagaggtgtctgaatgtg agtgtatgca tgctgcgtgt gtctgtgttt atgtttgttt 1740 gtttgtttggggcaagaaag attctaggac aagagctagg catgtacttc tgaccaggtg 1800 ggtaagcaactctaagtctg tatttgtatt ggtcattctc agtggaaatc ccttaggccc 1860 tctagtggttttcccctacc tgcatattgg ttttcatgtt ttatattcac tgttactatc 1920 ttctgtgtttaattaaaatt gttttctatc aaaaaaaaaa aaaaaaaaaa gggcggccgc 1980 34 1365 DNAHomo sapiens 34 atgtgtacaa agacaatccc agtcctctgg ggatgtttcc tcctgtggaatctctatgtc 60 tcatcctctc agaccattta ccctggaatc aaggcaagga ttactcagagggcacttgac 120 tatggtgttc aagctggaat gaagatgatt gagcaaatgc taaaagaaaagaaactccca 180 gatttaagcg gttctgagtc tcttgaattt ctaaaagttg attatgtaaactacaatttt 240 tcaaatataa aaatcagtgc cttttcattt ccaaatacct cattggcttttgtgcctgga 300 gtgggaatca aagcgctaac caaccatggc actgccaaca tcagcacagactgggggttc 360 gagtctccac tttttgttct gtataactcc tttgctgagc ccatggagaaacccatttta 420 aagaacttaa atgaaatgct ctgtcccatt attgcaagtg aagtcaaagcgctaaatgcc 480 aacctcagca cactggaggt tttaaccaag attgacaact acactctgctggattactcc 540 ctaatcagtt ctccagaaat tactgagaac taccttgacc tgaacttgaagggtgtattc 600 tacccactgg aaaacctcac cgaccccccc ttctcaccag ttccttttgtgctcccagaa 660 cgcagcaact ccatgctcta cattggaatc gccgagtatt tctttaaatctgcgtccttt 720 gctcatttca cagctggggt tttcaatctc actctctcca ccgaagagatttccaaccat 780 tttgttcaaa actctcaagg ccttggcaac gtgctctccc ggattgcagagatctacatc 840 ttgtcccagc ccttcatggt gaggatcatg gccacagagc ctcccataatcaatctacaa 900 ccaggcaatt tcaccctgga catccctgcc tccatcatga tgctcacccaacccaagaac 960 tccacagttg aaaccatcgt ttccatggac ttcgttgcta gtaccagtgttggcctggtt 1020 attttgggac aaagactggt ctgctccttg tctctgaaca gattccgccttgctttgcca 1080 gagtccaatc gcagcaacat tgaggtcttg aggtttgaaa atattctatcgtccattctt 1140 cactttggag tcctcccact ggccaatgca aaattgcagc aaggatttcctctgcccaat 1200 ccacacaaat tcttattcgt caattcagat attgaagttc ttgagggtttccttttgatt 1260 tccaccgacc tgaagtatga aacatcctca aagcagcagc caagtttccacgtatgggaa 1320 ggtctgaacc tgataagcag acagtggagg gggaagtcag cccct 136535 455 PRT Homo sapiens 35 Met Cys Thr Lys Thr Ile Pro Val Leu Trp GlyCys Phe Leu Leu Trp 1 5 10 15 Asn Leu Tyr Val Ser Ser Ser Gln Thr IleTyr Pro Gly Ile Lys Ala 20 25 30 Arg Ile Thr Gln Arg Ala Leu Asp Tyr GlyVal Gln Ala Gly Met Lys 35 40 45 Met Ile Glu Gln Met Leu Lys Glu Lys LysLeu Pro Asp Leu Ser Gly 50 55 60 Ser Glu Ser Leu Glu Phe Leu Lys Val AspTyr Val Asn Tyr Asn Phe 65 70 75 80 Ser Asn Ile Lys Ile Ser Ala Phe SerPhe Pro Asn Thr Ser Leu Ala 85 90 95 Phe Val Pro Gly Val Gly Ile Lys AlaLeu Thr Asn His Gly Thr Ala 100 105 110 Asn Ile Ser Thr Asp Trp Gly PheGlu Ser Pro Leu Phe Val Leu Tyr 115 120 125 Asn Ser Phe Ala Glu Pro MetGlu Lys Pro Ile Leu Lys Asn Leu Asn 130 135 140 Glu Met Leu Cys Pro IleIle Ala Ser Glu Val Lys Ala Leu Asn Ala 145 150 155 160 Asn Leu Ser ThrLeu Glu Val Leu Thr Lys Ile Asp Asn Tyr Thr Leu 165 170 175 Leu Asp TyrSer Leu Ile Ser Ser Pro Glu Ile Thr Glu Asn Tyr Leu 180 185 190 Asp LeuAsn Leu Lys Gly Val Phe Tyr Pro Leu Glu Asn Leu Thr Asp 195 200 205 ProPro Phe Ser Pro Val Pro Phe Val Leu Pro Glu Arg Ser Asn Ser 210 215 220Met Leu Tyr Ile Gly Ile Ala Glu Tyr Phe Phe Lys Ser Ala Ser Phe 225 230235 240 Ala His Phe Thr Ala Gly Val Phe Asn Leu Thr Leu Ser Thr Glu Glu245 250 255 Ile Ser Asn His Phe Val Gln Asn Ser Gln Gly Leu Gly Asn ValLeu 260 265 270 Ser Arg Ile Ala Glu Ile Tyr Ile Leu Ser Gln Pro Phe MetVal Arg 275 280 285 Ile Met Ala Thr Glu Pro Pro Ile Ile Asn Leu Gln ProGly Asn Phe 290 295 300 Thr Leu Asp Ile Pro Ala Ser Ile Met Met Leu ThrGln Pro Lys Asn 305 310 315 320 Ser Thr Val Glu Thr Ile Val Ser Met AspPhe Val Ala Ser Thr Ser 325 330 335 Val Gly Leu Val Ile Leu Gly Gln ArgLeu Val Cys Ser Leu Ser Leu 340 345 350 Asn Arg Phe Arg Leu Ala Leu ProGlu Ser Asn Arg Ser Asn Ile Glu 355 360 365 Val Leu Arg Phe Glu Asn IleLeu Ser Ser Ile Leu His Phe Gly Val 370 375 380 Leu Pro Leu Ala Asn AlaLys Leu Gln Gln Gly Phe Pro Leu Pro Asn 385 390 395 400 Pro His Lys PheLeu Phe Val Asn Ser Asp Ile Glu Val Leu Glu Gly 405 410 415 Phe Leu LeuIle Ser Thr Asp Leu Lys Tyr Glu Thr Ser Ser Lys Gln 420 425 430 Gln ProSer Phe His Val Trp Glu Gly Leu Asn Leu Ile Ser Arg Gln 435 440 445 TrpArg Gly Lys Ser Ala Pro 450 455 36 23 PRT Homo sapiens 36 Met Cys ThrLys Thr Ile Pro Val Leu Trp Gly Cys Phe Leu Leu Trp 1 5 10 15 Asn LeuTyr Val Ser Ser Ser 20 37 432 PRT Homo sapiens 37 Gln Thr Ile Tyr ProGly Ile Lys Ala Arg Ile Thr Gln Arg Ala Leu 1 5 10 15 Asp Tyr Gly ValGln Ala Gly Met Lys Met Ile Glu Gln Met Leu Lys 20 25 30 Glu Lys Lys LeuPro Asp Leu Ser Gly Ser Glu Ser Leu Glu Phe Leu 35 40 45 Lys Val Asp TyrVal Asn Tyr Asn Phe Ser Asn Ile Lys Ile Ser Ala 50 55 60 Phe Ser Phe ProAsn Thr Ser Leu Ala Phe Val Pro Gly Val Gly Ile 65 70 75 80 Lys Ala LeuThr Asn His Gly Thr Ala Asn Ile Ser Thr Asp Trp Gly 85 90 95 Phe Glu SerPro Leu Phe Val Leu Tyr Asn Ser Phe Ala Glu Pro Met 100 105 110 Glu LysPro Ile Leu Lys Asn Leu Asn Glu Met Leu Cys Pro Ile Ile 115 120 125 AlaSer Glu Val Lys Ala Leu Asn Ala Asn Leu Ser Thr Leu Glu Val 130 135 140Leu Thr Lys Ile Asp Asn Tyr Thr Leu Leu Asp Tyr Ser Leu Ile Ser 145 150155 160 Ser Pro Glu Ile Thr Glu Asn Tyr Leu Asp Leu Asn Leu Lys Gly Val165 170 175 Phe Tyr Pro Leu Glu Asn Leu Thr Asp Pro Pro Phe Ser Pro ValPro 180 185 190 Phe Val Leu Pro Glu Arg Ser Asn Ser Met Leu Tyr Ile GlyIle Ala 195 200 205 Glu Tyr Phe Phe Lys Ser Ala Ser Phe Ala His Phe ThrAla Gly Val 210 215 220 Phe Asn Leu Thr Leu Ser Thr Glu Glu Ile Ser AsnHis Phe Val Gln 225 230 235 240 Asn Ser Gln Gly Leu Gly Asn Val Leu SerArg Ile Ala Glu Ile Tyr 245 250 255 Ile Leu Ser Gln Pro Phe Met Val ArgIle Met Ala Thr Glu Pro Pro 260 265 270 Ile Ile Asn Leu Gln Pro Gly AsnPhe Thr Leu Asp Ile Pro Ala Ser 275 280 285 Ile Met Met Leu Thr Gln ProLys Asn Ser Thr Val Glu Thr Ile Val 290 295 300 Ser Met Asp Phe Val AlaSer Thr Ser Val Gly Leu Val Ile Leu Gly 305 310 315 320 Gln Arg Leu ValCys Ser Leu Ser Leu Asn Arg Phe Arg Leu Ala Leu 325 330 335 Pro Glu SerAsn Arg Ser Asn Ile Glu Val Leu Arg Phe Glu Asn Ile 340 345 350 Leu SerSer Ile Leu His Phe Gly Val Leu Pro Leu Ala Asn Ala Lys 355 360 365 LeuGln Gln Gly Phe Pro Leu Pro Asn Pro His Lys Phe Leu Phe Val 370 375 380Asn Ser Asp Ile Glu Val Leu Glu Gly Phe Leu Leu Ile Ser Thr Asp 385 390395 400 Leu Lys Tyr Glu Thr Ser Ser Lys Gln Gln Pro Ser Phe His Val Trp405 410 415 Glu Gly Leu Asn Leu Ile Ser Arg Gln Trp Arg Gly Lys Ser AlaPro 420 425 430 38 483 PRT Homo sapiens 38 Met Ala Arg Gly Pro Cys AsnAla Pro Arg Trp Val Ser Leu Met Val 1 5 10 15 Leu Val Ala Ile Gly ThrAla Val Thr Ala Ala Val Asn Pro Gly Val 20 25 30 Val Val Arg Ile Ser GlnLys Gly Leu Asp Tyr Ala Ser Gln Gln Gly 35 40 45 Thr Ala Ala Leu Gln LysGlu Leu Lys Arg Ile Lys Ile Pro Asp Tyr 50 55 60 Ser Asp Ser Phe Lys IleLys His Leu Gly Lys Gly His Tyr Ser Phe 65 70 75 80 Tyr Ser Met Asp IleArg Glu Phe Gln Leu Pro Ser Ser Gln Ile Ser 85 90 95 Met Val Pro Asn ValGly Leu Lys Phe Ser Ile Ser Asn Ala Asn Ile 100 105 110 Lys Ile Ser GlyLys Trp Lys Ala Gln Lys Arg Phe Leu Lys Met Ser 115 120 125 Gly Asn PheAsp Leu Ser Ile Glu Gly Met Ser Ile Ser Ala Asp Leu 130 135 140 Lys LeuGly Ser Asn Pro Thr Ser Gly Lys Pro Thr Ile Thr Cys Ser 145 150 155 160Ser Cys Ser Ser His Ile Asn Ser Val His Val His Ile Ser Lys Ser 165 170175 Lys Val Gly Trp Leu Ile Gln Leu Phe His Lys Lys Ile Glu Ser Ala 180185 190 Leu Arg Asn Lys Met Asn Ser Gln Val Cys Glu Lys Val Thr Asn Ser195 200 205 Val Ser Ser Lys Leu Gln Pro Tyr Phe Gln Thr Leu Pro Val MetThr 210 215 220 Lys Ile Asp Ser Val Ala Gly Ile Asn Tyr Gly Leu Val AlaPro Pro 225 230 235 240 Ala Thr Thr Ala Glu Thr Leu Asp Val Gln Met LysGly Glu Phe Tyr 245 250 255 Ser Glu Asn His His Asn Pro Pro Pro Phe AlaPro Pro Val Met Glu 260 265 270 Phe Pro Ala Ala His Asp Arg Met Val TyrLeu Gly Leu Ser Asp Tyr 275 280 285 Phe Phe Asn Thr Ala Gly Leu Val TyrGln Glu Ala Gly Val Leu Lys 290 295 300 Met Thr Leu Arg Asp Asp Met IlePro Lys Glu Ser Lys Phe Arg Leu 305 310 315 320 Thr Thr Lys Phe Phe GlyThr Phe Leu Pro Glu Val Ala Lys Lys Phe 325 330 335 Pro Asn Met Lys IleGln Ile His Val Ser Ala Ser Thr Pro Pro His 340 345 350 Leu Ser Val GlnPro Thr Gly Leu Thr Phe Tyr Pro Ala Val Asp Val 355 360 365 Gln Ala PheAla Val Leu Pro Asn Ser Ser Leu Ala Ser Leu Phe Leu 370 375 380 Ile GlyMet His Thr Thr Gly Ser Met Glu Val Ser Ala Glu Ser Asn 385 390 395 400Arg Leu Val Gly Glu Leu Lys Leu Asp Arg Leu Leu Leu Glu Leu Lys 405 410415 His Ser Asn Ile Gly Pro Phe Pro Val Glu Leu Leu Gln Asp Ile Met 420425 430 Asn Tyr Ile Val Pro Ile Leu Val Leu Pro Arg Val Asn Glu Lys Leu435 440 445 Gln Lys Gly Phe Pro Leu Pro Thr Pro Ala Arg Val Gln Leu TyrAsn 450 455 460 Val Val Leu Gln Pro His Gln Asn Phe Leu Leu Phe Gly AlaAsp Val 465 470 475 480 Val Tyr Lys 39 481 PRT Homo sapiens 39 Met GlyAla Leu Ala Arg Ala Leu Pro Ser Ile Leu Leu Ala Leu Leu 1 5 10 15 LeuThr Ser Thr Pro Glu Ala Leu Gly Ala Asn Pro Gly Leu Val Ala 20 25 30 ArgIle Thr Asp Lys Gly Leu Gln Tyr Ala Ala Gln Glu Gly Leu Leu 35 40 45 AlaLeu Gln Ser Glu Leu Leu Arg Ile Thr Leu Pro Asp Phe Thr Gly 50 55 60 AspLeu Arg Ile Pro His Val Gly Arg Gly Arg Tyr Glu Phe His Ser 65 70 75 80Leu Asn Ile His Glu Phe Gln Leu Pro Ser Ser Gln Ile Ser Met Val 85 90 95Pro Asn Val Gly Leu Lys Phe Ser Ile Ser Asn Ala Asn Ile Lys Ile 100 105110 Ser Gly Lys Trp Lys Ala Gln Lys Arg Phe Leu Lys Met Ser Gly Asn 115120 125 Phe Asp Leu Ser Ile Glu Gly Met Ser Ile Ser Ala Asp Leu Lys Leu130 135 140 Gly Ser Asn Pro Thr Ser Gly Lys Pro Thr Ile Thr Cys Ser SerCys 145 150 155 160 Ser Ser His Ile Asn Ser Val His Val His Ile Ser LysSer Lys Val 165 170 175 Gly Trp Leu Ile Gln Leu Phe His Lys Lys Ile GluSer Ala Leu Arg 180 185 190 Asn Lys Met Asn Ser Gln Val Cys Glu Lys ValThr Asn Ser Val Ser 195 200 205 Ser Lys Leu Gln Pro Tyr Phe Gln Thr LeuPro Val Met Thr Lys Ile 210 215 220 Asp Ser Val Ala Gly Ile Asn Tyr GlyLeu Val Ala Pro Pro Ala Thr 225 230 235 240 Thr Ala Glu Thr Leu Asp ValGln Met Lys Gly Glu Phe Tyr Ser Glu 245 250 255 Asn His His Asn Pro ProPro Phe Ala Pro Pro Val Met Glu Phe Pro 260 265 270 Ala Ala His Asp ArgMet Val Tyr Leu Gly Leu Ser Asp Tyr Phe Phe 275 280 285 Asn Thr Ala GlyLeu Val Tyr Gln Glu Ala Gly Val Leu Lys Met Thr 290 295 300 Leu Arg AspAsp Met Ile Pro Lys Glu Ser Lys Phe Arg Leu Thr Thr 305 310 315 320 LysPhe Phe Gly Thr Phe Leu Pro Glu Val Ala Lys Lys Phe Pro Asn 325 330 335Met Lys Ile Gln Ile His Val Ser Ala Ser Thr Pro Pro His Leu Ser 340 345350 Val Gln Pro Thr Gly Leu Thr Phe Tyr Pro Ala Val Asp Val Gln Ala 355360 365 Leu Ala Val Leu Pro Asn Ser Ser Leu Ala Ser Leu Phe Leu Ile Gly370 375 380 Met His Thr Thr Gly Ser Met Glu Val Ser Ala Glu Ser Asn ArgLeu 385 390 395 400 Val Gly Glu Leu Lys Leu Asp Arg Leu Leu Leu Glu LeuLys His Ser 405 410 415 Asn Ile Gly Pro Phe Pro Val Glu Leu Leu Gln AspIle Met Asn Tyr 420 425 430 Ile Val Pro Ile Leu Val Leu Pro Arg Val AsnGlu Lys Leu Gln Lys 435 440 445 Gly Phe Pro Leu Pro Thr Pro Ala Arg ValGln Leu Tyr Asn Val Val 450 455 460 Leu Gln Pro His Gln Asn Phe Leu LeuPhe Gly Ala Asp Val Val Tyr 465 470 475 480 Lys 40 383 PRTCaenorhabditis elegans 40 Met Arg Ile Ala His Ala Ser Ser Arg Gly AsnIle Ser Ile Phe Ser 1 5 10 15 Val Phe Leu Ile Pro Leu Ile Ala Tyr IleLeu Ile Leu Pro Gly Val 20 25 30 Arg Arg Lys Arg Val Val Thr Thr Val ThrTyr Val Leu Met Leu Ala 35 40 45 Val Gly Gly Ala Leu Ile Ala Ser Leu IleTyr Pro Cys Trp Ala Ser 50 55 60 Gly Ser Gln Met Ile Tyr Thr Gln Phe ArgGly His Ser Asn Glu Arg 65 70 75 80 Ile Leu Ala Lys Ile Gly Val Glu IleGly Leu Gln Lys Val Asn Val 85 90 95 Thr Leu Lys Phe Glu Arg Leu Leu SerSer Asn Asp Val Leu Pro Gly 100 105 110 Ser Asp Met Thr Glu Leu Tyr TyrAsn Glu Gly Phe Asp Ile Ser Gly 115 120 125 Ile Ser Ser Met Ala Glu AlaLeu His His Gly Leu Glu Asn Gly Leu 130 135 140 Pro Tyr Pro Met Leu SerVal Leu Glu Tyr Phe Ser Leu Asn Gln Asp 145 150 155 160 Ser Phe Asp TrpGly Arg His Tyr Arg Val Ala Gly His Tyr Thr His 165 170 175 Ala Ala IleTrp Phe Ala Phe Ala Cys Trp Cys Leu Ser Val Val Leu 180 185 190 Met LeuPhe Leu Pro His Asn Ala Tyr Lys Ser Ile Leu Ala Thr Gly 195 200 205 IleSer Cys Leu Ile Ala Cys Leu Val Tyr Leu Leu Leu Ser Pro Cys 210 215 220Glu Leu Arg Ile Ala Phe Thr Gly Glu Asn Phe Glu Arg Val Asp Leu 225 230235 240 Thr Ala Thr Phe Ser Phe Cys Phe Tyr Leu Ile Phe Ala Ile Gly Ile245 250 255 Leu Cys Val Leu Cys Gly Leu Gly Leu Gly Ile Cys Glu His TrpArg 260 265 270 Ile Tyr Thr Leu Ser Thr Phe Leu Asp Ala Ser Leu Asp GluHis Val 275 280 285 Gly Pro Lys Trp Lys Lys Leu Pro Thr Gly Gly Pro AlaLeu Gln Gly 290 295 300 Val Gln Ile Gly Ala Tyr Gly Thr Asn Thr Thr AsnSer Ser Arg Asp 305 310 315 320 Lys Asn Asp Ile Ser Ser Asp Lys Thr AlaGly Ser Ser Gly Phe Gln 325 330 335 Ser Arg Thr Ser Thr Cys Gln Ser SerAla Ser Ser Ala Ser Leu Arg 340 345 350 Ser Gln Ser Ser Ile Glu Thr ValHis Asp Glu Ala Glu Leu Glu Arg 355 360 365 Thr His Val His Phe Leu GlnGlu Pro Cys Ser Ser Ser Ser Thr 370 375 380 41 399 PRT Homo sapiens 41Met Lys Met Arg Phe Leu Gly Leu Val Val Cys Leu Val Leu Trp Pro 1 5 1015 Leu His Ser Glu Gly Ser Gly Gly Lys Leu Thr Ala Val Asp Pro Glu 20 2530 Thr Asn Met Asn Val Ser Glu Ile Ile Ser Tyr Trp Gly Phe Pro Ser 35 4045 Glu Glu Tyr Leu Val Glu Thr Glu Asp Gly Tyr Ile Leu Cys Leu Asn 50 5560 Arg Ile Pro His Gly Arg Lys Asn His Ser Asp Lys Gly Pro Lys Pro 65 7075 80 Val Val Phe Leu Gln His Gly Leu Leu Ala Asp Ser Ser Asn Trp Val 8590 95 Thr Asn Leu Ala Asn Ser Ser Leu Gly Phe Ile Leu Ala Asp Ala Gly100 105 110 Phe Asp Val Trp Met Gly Asn Ser Arg Gly Asn Thr Trp Ser ArgLys 115 120 125 His Lys Thr Leu Ser Val Ser Gln Asp Glu Phe Trp Ala PheSer Tyr 130 135 140 Asp Glu Met Ala Lys Tyr Asp Leu Pro Ala Ser Ile AsnPhe Ile Leu 145 150 155 160 Asn Lys Thr Gly Gln Glu Gln Val Tyr Tyr ValGly His Ser Gln Gly 165 170 175 Thr Thr Ile Gly Phe Ile Ala Phe Ser GlnIle Pro Glu Leu Ala Lys 180 185 190 Arg Ile Lys Met Phe Phe Ala Leu GlyPro Val Ala Ser Val Ala Phe 195 200 205 Cys Thr Ser Pro Met Ala Lys LeuGly Arg Leu Pro Asp His Leu Ile 210 215 220 Lys Asp Leu Phe Gly Asp LysGlu Phe Leu Pro Gln Ser Ala Phe Leu 225 230 235 240 Lys Trp Leu Gly ThrHis Val Cys Thr His Val Ile Leu Lys Glu Leu 245 250 255 Cys Gly Asn LeuCys Phe Leu Leu Cys Gly Phe Asn Glu Arg Asn Leu 260 265 270 Asn Met SerArg Val Asp Val Tyr Thr Thr His Ser Pro Ala Gly Thr 275 280 285 Ser ValGln Asn Met Leu His Trp Ser Gln Ala Val Lys Phe Gln Lys 290 295 300 PheGln Ala Phe Asp Trp Gly Ser Ser Ala Lys Asn Tyr Phe His Tyr 305 310 315320 Asn Gln Ser Tyr Pro Pro Thr Tyr Asn Val Lys Asp Met Leu Val Pro 325330 335 Thr Ala Val Trp Ser Gly Gly His Asp Trp Leu Ala Asp Val Tyr Asp340 345 350 Val Asn Ile Leu Leu Thr Gln Ile Thr Asn Leu Val Phe His GluSer 355 360 365 Ile Pro Glu Trp Glu His Leu Asp Phe Ile Trp Gly Leu AspAla Pro 370 375 380 Trp Arg Leu Tyr Asn Lys Ile Ile Asn Leu Met Arg LysTyr Gln 385 390 395 42 19 PRT Mus sp. 42 Met Ala Pro Pro Ala Ala Arg LeuAla Leu Leu Ser Ala Ala Ala Leu 1 5 10 15 Thr Leu Ala 43 451 PRT Mus sp.43 Ala Arg Pro Ala Pro Gly Pro Arg Ser Gly Pro Glu Cys Phe Thr Ala 1 510 15 Asn Gly Ala Asp Tyr Arg Gly Thr Gln Ser Trp Thr Ala Leu Gln Gly 2025 30 Gly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe Gln His Pro Tyr Asn 3540 45 Thr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu Gly Glu His Asn Tyr 5055 60 Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp Cys Tyr Val Ala Glu 6570 75 80 His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu Ile Pro Ala Cys Gln85 90 95 Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His Gly Asn Pro Pro Pro100 105 110 Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu Thr Ile Gln ThrCys 115 120 125 Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe Ala Gly MetGlu Ser 130 135 140 Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro Asp Tyr TrpLys His Gly 145 150 155 160 Glu Ala Ala Ser Thr Glu Cys Asn Ser Val CysPhe Gly Asp His Thr 165 170 175 Gln Pro Cys Gly Gly Asp Gly Arg Ile IleLeu Phe Asp Thr Leu Val 180 185 190 Gly Ala Cys Gly Gly Asn Tyr Ser AlaMet Ala Ala Val Val Tyr Ser 195 200 205 Pro Asp Phe Pro Asp Thr Tyr AlaThr Gly Arg Val Cys Tyr Trp Thr 210 215 220 Ile Arg Val Pro Gly Ala SerArg Ile His Phe Asn Phe Thr Leu Phe 225 230 235 240 Asp Ile Arg Asp SerAla Asp Met Val Glu Leu Leu Asp Gly Tyr Thr 245 250 255 His Arg Val LeuVal Arg Leu Ser Gly Arg Ser Arg Pro Pro Leu Ser 260 265 270 Phe Asn ValSer Leu Asp Phe Val Ile Leu Tyr Phe Phe Ser Asp Arg 275 280 285 Ile AsnGln Ala Gln Gly Phe Ala Val Leu Tyr Gln Ala Thr Lys Glu 290 295 300 GluPro Pro Gln Glu Arg Pro Ala Val Asn Gln Thr Leu Ala Glu Val 305 310 315320 Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala Ala His Ser Ser Lys 325330 335 Val Leu Tyr Val Ile Thr Pro Ser Pro Ser His Pro Pro Gln Thr Ala340 345 350 Gln Val Ala Ile Pro Gly His Arg Gln Leu Gly Pro Thr Ala ThrGlu 355 360 365 Trp Lys Asp Gly Leu Cys Thr Ala Trp Arg Pro Ser Ser SerSer Gln 370 375 380 Ser Gln Gln Leu Ser Gln Arg Phe Phe Cys Met Ser HisLeu Asn Leu 385 390 395 400 Ile Glu Ser Leu His Gln Glu Thr Leu Gly ThrVal Val Ser Leu Gly 405 410 415 Leu Leu Glu Ile Ser Gly Pro Phe Ser MetAsn Leu Pro Leu Gln Ser 420 425 430 Pro Ser Leu Arg Arg Ser Ser Arg ValArg Val Asn Lys Met Thr Ala 435 440 445 Ile Pro Ser 450 44 150 PRT Mussp. 44 Lys Lys His Cys Trp Tyr Phe Glu Gly Leu Tyr Pro Thr Tyr Tyr Ile 15 10 15 Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser Arg Cys Cys Val Arg Ala20 25 30 Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp Phe Leu Leu Met Met Gly35 40 45 Val Leu Phe Cys Cys Gly Ala Gly Phe Phe Ile Arg Arg Arg Met Tyr50 55 60 Pro Pro Pro Leu Ile Glu Glu Pro Thr Phe Asn Val Ser Tyr Thr Arg65 70 75 80 Gln Pro Pro Asn Pro Ala Pro Gly Ala Gln Gln Met Gly Pro ProTyr 85 90 95 Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn Pro Val Gly Asn ThrMet 100 105 110 Ala Met Ala Phe Gln Val Gln Pro Asn Ser Pro His Gly GlyThr Thr 115 120 125 Tyr Pro Pro Pro Pro Ser Tyr Cys Asn Thr Pro Pro ProPro Tyr Glu 130 135 140 Gln Val Val Lys Asp Lys 145 150 45 2044 DNA Homosapiens 45 gtcgacccac gcgtccgggg aattgcagca ggaaaatatg tgaagagtttttaaacccac 60 aaattcttct tactttagaa ttagttgtta cattggcagg aaaaaataaatgcagatgtt 120 ggaccatgtt ggaaaccttg tcaagacagt ggattgtctc acacagaatggaaatgtggc 180 ttctgattct ggtggcgtat atgttccaga gaaatgtgaa ttcagtacatatgccaacta 240 aagctgtgga cccagaagca ttcatgaata ttagtgaaat catccaacatcaaggctatc 300 cctgtgagga atatgaagtc gcaactgaag atgggtatat cctttctgttaacaggattc 360 ctcgaggcct agtgcaacct aagaagacag gttccaggcc tgtggtgttactgcagcatg 420 gcctagttgg aggtgctagc aactggattt ccaacctgcc caacaatagcctgggcttca 480 ttctggcaga tgctggtttt gacgtgtgga tggggaacag caggggaaacgcctggtctc 540 gaaaacacaa gacactctcc atagaccaag atgagttctg ggctttcagttatgatgaga 600 tggctaggtt tgaccttcct gcagtgataa actttatttt gcagaaaacgggccaggaaa 660 agatctatta tgtcggctat tcacagggca ccaccatggg ctttattgcattttccacca 720 tgccagagct ggctcagaaa atcaaaatgt attttgcttt agcacccatagccactgtta 780 agcatgcaaa aagccccggg accaaatttt tgttgctgcc agatatgatgatcaagggat 840 tgtttggcaa aaaagaattt ctgtatcaga ccagatttct cagacaacttgttatttacc 900 tttgtggcca ggtgattctt gatcagattt gtagtaatat catgttacttctgggtggat 960 tcaacaccaa caatatgaac atgagccgag caagtgtata tgctgcccacactcttgctg 1020 gaacatctgt gcaaaatatt ctacactgga gccaggcagt gaattctggtgaactccggg 1080 catttgactg ggggagtgag accaaaaatc tggaaaaatg caatcagccaactcctgtaa 1140 ggtacagagt cagagatatg acggtcccta cagcaatgtg gacaggaggtcaggactggc 1200 tttcaaatcc agaagacgtg aaaatgctgc tctctgaggt gaccaacctcatctaccata 1260 agaatattcc tgaatgggct cacgtggatt tcatctgggg tttggatgctcctcaccgta 1320 tgtacaatga aatcatccat ctgatgcagc aggaggagac caacctttcccagggacggt 1380 gtgaggccgt attgtgaagc atctgacact gacgatctta ggacaacctcctgagggatg 1440 gggctaggac ccatgaaggc agaattacgg agagcagaga cctagtatacatttttcaga 1500 ttccctgcac ttggcactaa atccgacact tacatttaca ttttttttctgtaaattaaa 1560 gtacttatta ggtaaataga ggttttgtat gctattatat attctaccatcttgaagggt 1620 aggttttacc tgatagccag aaaatatcta gacattctct atatcattcaggtaaatctc 1680 tttaaaacac ctattgtttt ttctataagc catatttttg gagcactaaagtaaaatggc 1740 aaattgggac agatattgag gtctggagtc tgtggattat tgttgactttgacaaaataa 1800 gctagacatt ttcaccttgt tgccacagag acataacact acctcaggaagctgagctgc 1860 tttaaggaca acaacaacaa aatcagtgtt acagtatgga tgaaatctatgttaagcatt 1920 ctcagaataa ggccaagttt tatagttgca tctcagggaa gaaaattttataggatgttt 1980 atgagttctc caataaatgc attctgcatt acataaaaaa aaaaaaaaaaaaaagggcgg 2040 ccgc 2044 46 1269 DNA Homo sapiens 46 atgttggaaaccttgtcaag acagtggatt gtctcacaca gaatggaaat gtggcttctg 60 attctggtggcgtatatgtt ccagagaaat gtgaattcag tacatatgcc aactaaagct 120 gtggacccagaagcattcat gaatattagt gaaatcatcc aacatcaagg ctatccctgt 180 gaggaatatgaagtcgcaac tgaagatggg tatatccttt ctgttaacag gattcctcga 240 ggcctagtgcaacctaagaa gacaggttcc aggcctgtgg tgttactgca gcatggccta 300 gttggaggtgctagcaactg gatttccaac ctgcccaaca atagcctggg cttcattctg 360 gcagatgctggttttgacgt gtggatgggg aacagcaggg gaaacgcctg gtctcgaaaa 420 cacaagacactctccataga ccaagatgag ttctgggctt tcagttatga tgagatggct 480 aggtttgaccttcctgcagt gataaacttt attttgcaga aaacgggcca ggaaaagatc 540 tattatgtcggctattcaca gggcaccacc atgggcttta ttgcattttc caccatgcca 600 gagctggctcagaaaatcaa aatgtatttt gctttagcac ccatagccac tgttaagcat 660 gcaaaaagccccgggaccaa atttttgttg ctgccagata tgatgatcaa gggattgttt 720 ggcaaaaaagaatttctgta tcagaccaga tttctcagac aacttgttat ttacctttgt 780 ggccaggtgattcttgatca gatttgtagt aatatcatgt tacttctggg tggattcaac 840 accaacaatatgaacatgag ccgagcaagt gtatatgctg cccacactct tgctggaaca 900 tctgtgcaaaatattctaca ctggagccag gcagtgaatt ctggtgaact ccgggcattt 960 gactgggggagtgagaccaa aaatctggaa aaatgcaatc agccaactcc tgtaaggtac 1020 agagtcagagatatgacggt ccctacagca atgtggacag gaggtcagga ctggctttca 1080 aatccagaagacgtgaaaat gctgctctct gaggtgacca acctcatcta ccataagaat 1140 attcctgaatgggctcacgt ggatttcatc tggggtttgg atgctcctca ccgtatgtac 1200 aatgaaatcatccatctgat gcagcaggag gagaccaacc tttcccaggg acggtgtgag 1260 gccgtattg1269 47 423 PRT Homo sapiens 47 Met Leu Glu Thr Leu Ser Arg Gln Trp IleVal Ser His Arg Met Glu 1 5 10 15 Met Trp Leu Leu Ile Leu Val Ala TyrMet Phe Gln Arg Asn Val Asn 20 25 30 Ser Val His Met Pro Thr Lys Ala ValAsp Pro Glu Ala Phe Met Asn 35 40 45 Ile Ser Glu Ile Ile Gln His Gln GlyTyr Pro Cys Glu Glu Tyr Glu 50 55 60 Val Ala Thr Glu Asp Gly Tyr Ile LeuSer Val Asn Arg Ile Pro Arg 65 70 75 80 Gly Leu Val Gln Pro Lys Lys ThrGly Ser Arg Pro Val Val Leu Leu 85 90 95 Gln His Gly Leu Val Gly Gly AlaSer Asn Trp Ile Ser Asn Leu Pro 100 105 110 Asn Asn Ser Leu Gly Phe IleLeu Ala Asp Ala Gly Phe Asp Val Trp 115 120 125 Met Gly Asn Ser Arg GlyAsn Ala Trp Ser Arg Lys His Lys Thr Leu 130 135 140 Ser Ile Asp Gln AspGlu Phe Trp Ala Phe Ser Tyr Asp Glu Met Ala 145 150 155 160 Arg Phe AspLeu Pro Ala Val Ile Asn Phe Ile Leu Gln Lys Thr Gly 165 170 175 Gln GluLys Ile Tyr Tyr Val Gly Tyr Ser Gln Gly Thr Thr Met Gly 180 185 190 PheIle Ala Phe Ser Thr Met Pro Glu Leu Ala Gln Lys Ile Lys Met 195 200 205Tyr Phe Ala Leu Ala Pro Ile Ala Thr Val Lys His Ala Lys Ser Pro 210 215220 Gly Thr Lys Phe Leu Leu Leu Pro Asp Met Met Ile Lys Gly Leu Phe 225230 235 240 Gly Lys Lys Glu Phe Leu Tyr Gln Thr Arg Phe Leu Arg Gln LeuVal 245 250 255 Ile Tyr Leu Cys Gly Gln Val Ile Leu Asp Gln Ile Cys SerAsn Ile 260 265 270 Met Leu Leu Leu Gly Gly Phe Asn Thr Asn Asn Met AsnMet Ser Arg 275 280 285 Ala Ser Val Tyr Ala Ala His Thr Leu Ala Gly ThrSer Val Gln Asn 290 295 300 Ile Leu His Trp Ser Gln Ala Val Asn Ser GlyGlu Leu Arg Ala Phe 305 310 315 320 Asp Trp Gly Ser Glu Thr Lys Asn LeuGlu Lys Cys Asn Gln Pro Thr 325 330 335 Pro Val Arg Tyr Arg Val Arg AspMet Thr Val Pro Thr Ala Met Trp 340 345 350 Thr Gly Gly Gln Asp Trp LeuSer Asn Pro Glu Asp Val Lys Met Leu 355 360 365 Leu Ser Glu Val Thr AsnLeu Ile Tyr His Lys Asn Ile Pro Glu Trp 370 375 380 Ala His Val Asp PheIle Trp Gly Leu Asp Ala Pro His Arg Met Tyr 385 390 395 400 Asn Glu IleIle His Leu Met Gln Gln Glu Glu Thr Asn Leu Ser Gln 405 410 415 Gly ArgCys Glu Ala Val Leu 420 48 33 PRT Homo sapiens 48 Met Leu Glu Thr LeuSer Arg Gln Trp Ile Val Ser His Arg Met Glu 1 5 10 15 Met Trp Leu LeuIle Leu Val Ala Tyr Met Phe Gln Arg Asn Val Asn 20 25 30 Ser 49 390 PRTHomo sapiens 49 Val His Met Pro Thr Lys Ala Val Asp Pro Glu Ala Phe MetAsn Ile 1 5 10 15 Ser Glu Ile Ile Gln His Gln Gly Tyr Pro Cys Glu GluTyr Glu Val 20 25 30 Ala Thr Glu Asp Gly Tyr Ile Leu Ser Val Asn Arg IlePro Arg Gly 35 40 45 Leu Val Gln Pro Lys Lys Thr Gly Ser Arg Pro Val ValLeu Leu Gln 50 55 60 His Gly Leu Val Gly Gly Ala Ser Asn Trp Ile Ser AsnLeu Pro Asn 65 70 75 80 Asn Ser Leu Gly Phe Ile Leu Ala Asp Ala Gly PheAsp Val Trp Met 85 90 95 Gly Asn Ser Arg Gly Asn Ala Trp Ser Arg Lys HisLys Thr Leu Ser 100 105 110 Ile Asp Gln Asp Glu Phe Trp Ala Phe Ser TyrAsp Glu Met Ala Arg 115 120 125 Phe Asp Leu Pro Ala Val Ile Asn Phe IleLeu Gln Lys Thr Gly Gln 130 135 140 Glu Lys Ile Tyr Tyr Val Gly Tyr SerGln Gly Thr Thr Met Gly Phe 145 150 155 160 Ile Ala Phe Ser Thr Met ProGlu Leu Ala Gln Lys Ile Lys Met Tyr 165 170 175 Phe Ala Leu Ala Pro IleAla Thr Val Lys His Ala Lys Ser Pro Gly 180 185 190 Thr Lys Phe Leu LeuLeu Pro Asp Met Met Ile Lys Gly Leu Phe Gly 195 200 205 Lys Lys Glu PheLeu Tyr Gln Thr Arg Phe Leu Arg Gln Leu Val Ile 210 215 220 Tyr Leu CysGly Gln Val Ile Leu Asp Gln Ile Cys Ser Asn Ile Met 225 230 235 240 LeuLeu Leu Gly Gly Phe Asn Thr Asn Asn Met Asn Met Ser Arg Ala 245 250 255Ser Val Tyr Ala Ala His Thr Leu Ala Gly Thr Ser Val Gln Asn Ile 260 265270 Leu His Trp Ser Gln Ala Val Asn Ser Gly Glu Leu Arg Ala Phe Asp 275280 285 Trp Gly Ser Glu Thr Lys Asn Leu Glu Lys Cys Asn Gln Pro Thr Pro290 295 300 Val Arg Tyr Arg Val Arg Asp Met Thr Val Pro Thr Ala Met TrpThr 305 310 315 320 Gly Gly Gln Asp Trp Leu Ser Asn Pro Glu Asp Val LysMet Leu Leu 325 330 335 Ser Glu Val Thr Asn Leu Ile Tyr His Lys Asn IlePro Glu Trp Ala 340 345 350 His Val Asp Phe Ile Trp Gly Leu Asp Ala ProHis Arg Met Tyr Asn 355 360 365 Glu Ile Ile His Leu Met Gln Gln Glu GluThr Asn Leu Ser Gln Gly 370 375 380 Arg Cys Glu Ala Val Leu 385 390 50221 PRT Homo sapiens 50 Val His Met Pro Thr Lys Ala Val Asp Pro Glu AlaPhe Met Asn Ile 1 5 10 15 Ser Glu Ile Ile Gln His Gln Gly Tyr Pro CysGlu Glu Tyr Glu Val 20 25 30 Ala Thr Glu Asp Gly Tyr Ile Leu Ser Val AsnArg Ile Pro Arg Gly 35 40 45 Leu Val Gln Pro Lys Lys Thr Gly Ser Arg ProVal Val Leu Leu Gln 50 55 60 His Gly Leu Val Gly Gly Ala Ser Asn Trp IleSer Asn Leu Pro Asn 65 70 75 80 Asn Ser Leu Gly Phe Ile Leu Ala Asp AlaGly Phe Asp Val Trp Met 85 90 95 Gly Asn Ser Arg Gly Asn Ala Trp Ser ArgLys His Lys Thr Leu Ser 100 105 110 Ile Asp Gln Asp Glu Phe Trp Ala PheSer Tyr Asp Glu Met Ala Arg 115 120 125 Phe Asp Leu Pro Ala Val Ile AsnPhe Ile Leu Gln Lys Thr Gly Gln 130 135 140 Glu Lys Ile Tyr Tyr Val GlyTyr Ser Gln Gly Thr Thr Met Gly Phe 145 150 155 160 Ile Ala Phe Ser ThrMet Pro Glu Leu Ala Gln Lys Ile Lys Met Tyr 165 170 175 Phe Ala Leu AlaPro Ile Ala Thr Val Lys His Ala Lys Ser Pro Gly 180 185 190 Thr Lys PheLeu Leu Leu Pro Asp Met Met Ile Lys Gly Leu Phe Gly 195 200 205 Lys LysGlu Phe Leu Tyr Gln Thr Arg Phe Leu Arg Gln 210 215 220 51 25 PRT Homosapiens 51 Leu Val Ile Tyr Leu Cys Gly Gln Val Ile Leu Asp Gln Ile CysSer 1 5 10 15 Asn Ile Met Leu Leu Leu Gly Gly Phe 20 25 52 144 PRT Homosapiens 52 Asn Thr Asn Asn Met Asn Met Ser Arg Ala Ser Val Tyr Ala AlaHis 1 5 10 15 Thr Leu Ala Gly Thr Ser Val Gln Asn Ile Leu His Trp SerGln Ala 20 25 30 Val Asn Ser Gly Glu Leu Arg Ala Phe Asp Trp Gly Ser GluThr Lys 35 40 45 Asn Leu Glu Lys Cys Asn Gln Pro Thr Pro Val Arg Tyr ArgVal Arg 50 55 60 Asp Met Thr Val Pro Thr Ala Met Trp Thr Gly Gly Gln AspTrp Leu 65 70 75 80 Ser Asn Pro Glu Asp Val Lys Met Leu Leu Ser Glu ValThr Asn Leu 85 90 95 Ile Tyr His Lys Asn Ile Pro Glu Trp Ala His Val AspPhe Ile Trp 100 105 110 Gly Leu Asp Ala Pro His Arg Met Tyr Asn Glu IleIle His Leu Met 115 120 125 Gln Gln Glu Glu Thr Asn Leu Ser Gln Gly ArgCys Glu Ala Val Leu 130 135 140 53 2133 DNA Homo sapiens 53 gtcgacccacgcgtccacgg cgagggctcc cggggcgcag cattgccccc cctgcaccac 60 ctcaccaagatggctacttt gggacacaca ttccccttct atgctggccc caagccaacc 120 ttcccgatggacaccacttt ggccagcatc atcatgatct ttctgactgc actggccacg 180 ttcatcgtcatcctgcctgg cattcgggga aagacgaggc tgttctggct gcttcgggtg 240 gtgaccagcttattcatcgg ggctgcaatc ctggctgtga atttcagttc tgagtggtct 300 gtgggccaggtcagcaccaa cacatcatac aaggccttca gttctgagtg gatcagcgct 360 gatattgggctgcaggtcgg gctgggtgga gtcaacatca cactcacagg gacccccgtg 420 cagcagctgaatgagaccat caattacaac gaggagttca cctggcgcct gggtgagaac 480 tatgctgaggagtgtgcaaa ggctctggag aaggggctgc cagaccctgt gttgtaccta 540 gctgagaagttcactccaag aagcccatgt ggcctatacc gccagtaccg cctggcggga 600 cactacacctcagccatgct atgggtggca ttcctctgct ggctgctggc caatgtgatg 660 ctctccatgcctgtgctggt atatggtggc tacatgctat tggccacggg catcttccag 720 ctgttggctctgctcttctt ctccatggcc acatcactca cctcaccctg tcccctgcac 780 ctgggcgcttctgtgctgca tactcaccat gggcctgcct tctggatcac attgaccaca 840 ggactgctgtgtgtgctgct gggcctggct atggcggtgg cccacaggat gcagcctcac 900 aggctgaaggctttcttcaa ccagagtgtg gatgaagacc ccatgctgga gtggagtcct 960 gaggaaggtggactcctgag cccccgctac cggtccatgg ctgacagtcc caagtcccag 1020 gacattcccctgtcagaggc ttcctccacc aaggcatact gtaaggaggc acaccccaaa 1080 gatcctgattgtgctttata acattcctcc ccgtggaggc cacctggact tccagtctgg 1140 ctccaaacctcattggcgcc ccataaaacc agcagaactg ccctcagggt ggctgttacc 1200 agacacccagcaccaatcta cagacggagt agaaaaagga ggctctatat actgatgtta 1260 aaaaacaaaacaaaacaaaa agccctaagg gactgaagag atgctgggcc tgtccataaa 1320 gcctgttgccatgataaggc caagcagggg ctagcttatc tgcacagcaa cccagccttt 1380 ccgtgctgccttgcctcttc aagatgctat tcactgaaac ctaacttcac ccccataaca 1440 ccagcagggtgggggttaca tatgattctc ctatggtttc ctctcatccc tcggcacctc 1500 ttgttttcctttttcctggg ttccttttgt tcttccttta cttctccagc ttgtgtggcc 1560 ttttggtacaatgaaagaca gcactggaaa ggaggggaaa ccaaacttct catcctaggt 1620 ctaacattaaccaactatgc cacattctct ttgagcttca gttcccaaat ttgctacata 1680 agattgcaagacttgccaag aatcttggga tttatctttc tatgccttgc tgacacctac 1740 cttggccctcaaacaccacc tcacaagaag ccaggtggga agttagggaa tcaactccaa 1800 aacgctattccttcccaccc cactcagctg ggctagctga gtggcatcca ggacggggga 1860 gtgggtgacctgcctcatca ctgccaccta acgtccccct ggggtggttc agaaagatgc 1920 tagctctggtagggtccctc cggcctcact agagggcgcc cctattactc tggagtcgac 1980 gcagagaatcaggtttcaca gcactgcgga gagtgtacta ggctgtctcc agcccagcga 2040 agctcatgaggacgtgcgac cccggcgcgg agaagccatg aaaattaatg ggaaaaacag 2100 tttttaaaaaaaaaaaaaaa aaagggcggc cgc 2133 54 1029 DNA Homo sapiens 54 atggctactttgggacacac attccccttc tatgctggcc ccaagccaac cttcccgatg 60 gacaccactttggccagcat catcatgatc tttctgactg cactggccac gttcatcgtc 120 atcctgcctggcattcgggg aaagacgagg ctgttctggc tgcttcgggt ggtgaccagc 180 ttattcatcggggctgcaat cctggctgtg aatttcagtt ctgagtggtc tgtgggccag 240 gtcagcaccaacacatcata caaggccttc agttctgagt ggatcagcgc tgatattggg 300 ctgcaggtcgggctgggtgg agtcaacatc acactcacag ggacccccgt gcagcagctg 360 aatgagaccatcaattacaa cgaggagttc acctggcgcc tgggtgagaa ctatgctgag 420 gagtgtgcaaaggctctgga gaaggggctg ccagaccctg tgttgtacct agctgagaag 480 ttcactccaagaagcccatg tggcctatac cgccagtacc gcctggcggg acactacacc 540 tcagccatgctatgggtggc attcctctgc tggctgctgg ccaatgtgat gctctccatg 600 cctgtgctggtatatggtgg ctacatgcta ttggccacgg gcatcttcca gctgttggct 660 ctgctcttcttctccatggc cacatcactc acctcaccct gtcccctgca cctgggcgct 720 tctgtgctgcatactcacca tgggcctgcc ttctggatca cattgaccac aggactgctg 780 tgtgtgctgctgggcctggc tatggcggtg gcccacagga tgcagcctca caggctgaag 840 gctttcttcaaccagagtgt ggatgaagac cccatgctgg agtggagtcc tgaggaaggt 900 ggactcctgagcccccgcta ccggtccatg gctgacagtc ccaagtccca ggacattccc 960 ctgtcagaggcttcctccac caaggcatac tgtaaggagg cacaccccaa agatcctgat 1020 tgtgcttta1029 55 343 PRT Homo sapiens 55 Met Ala Thr Leu Gly His Thr Phe Pro PheTyr Ala Gly Pro Lys Pro 1 5 10 15 Thr Phe Pro Met Asp Thr Thr Leu AlaSer Ile Ile Met Ile Phe Leu 20 25 30 Thr Ala Leu Ala Thr Phe Ile Val IleLeu Pro Gly Ile Arg Gly Lys 35 40 45 Thr Arg Leu Phe Trp Leu Leu Arg ValVal Thr Ser Leu Phe Ile Gly 50 55 60 Ala Ala Ile Leu Ala Val Asn Phe SerSer Glu Trp Ser Val Gly Gln 65 70 75 80 Val Ser Thr Asn Thr Ser Tyr LysAla Phe Ser Ser Glu Trp Ile Ser 85 90 95 Ala Asp Ile Gly Leu Gln Val GlyLeu Gly Gly Val Asn Ile Thr Leu 100 105 110 Thr Gly Thr Pro Val Gln GlnLeu Asn Glu Thr Ile Asn Tyr Asn Glu 115 120 125 Glu Phe Thr Trp Arg LeuGly Glu Asn Tyr Ala Glu Glu Cys Ala Lys 130 135 140 Ala Leu Glu Lys GlyLeu Pro Asp Pro Val Leu Tyr Leu Ala Glu Lys 145 150 155 160 Phe Thr ProArg Ser Pro Cys Gly Leu Tyr Arg Gln Tyr Arg Leu Ala 165 170 175 Gly HisTyr Thr Ser Ala Met Leu Trp Val Ala Phe Leu Cys Trp Leu 180 185 190 LeuAla Asn Val Met Leu Ser Met Pro Val Leu Val Tyr Gly Gly Tyr 195 200 205Met Leu Leu Ala Thr Gly Ile Phe Gln Leu Leu Ala Leu Leu Phe Phe 210 215220 Ser Met Ala Thr Ser Leu Thr Ser Pro Cys Pro Leu His Leu Gly Ala 225230 235 240 Ser Val Leu His Thr His His Gly Pro Ala Phe Trp Ile Thr LeuThr 245 250 255 Thr Gly Leu Leu Cys Val Leu Leu Gly Leu Ala Met Ala ValAla His 260 265 270 Arg Met Gln Pro His Arg Leu Lys Ala Phe Phe Asn GlnSer Val Asp 275 280 285 Glu Asp Pro Met Leu Glu Trp Ser Pro Glu Glu GlyGly Leu Leu Ser 290 295 300 Pro Arg Tyr Arg Ser Met Ala Asp Ser Pro LysSer Gln Asp Ile Pro 305 310 315 320 Leu Ser Glu Ala Ser Ser Thr Lys AlaTyr Cys Lys Glu Ala His Pro 325 330 335 Lys Asp Pro Asp Cys Ala Leu 34056 23 PRT Homo sapiens 56 Met Ala Thr Leu Gly His Thr Phe Pro Phe TyrAla Gly Pro Lys Pro 1 5 10 15 Thr Phe Pro Met Asp Thr Thr 20 57 112 PRTHomo sapiens 57 Asn Phe Ser Ser Glu Trp Ser Val Gly Gln Val Ser Thr AsnThr Ser 1 5 10 15 Tyr Lys Ala Phe Ser Ser Glu Trp Ile Ser Ala Asp IleGly Leu Gln 20 25 30 Val Gly Leu Gly Gly Val Asn Ile Thr Leu Thr Gly ThrPro Val Gln 35 40 45 Gln Leu Asn Glu Thr Ile Asn Tyr Asn Glu Glu Phe ThrTrp Arg Leu 50 55 60 Gly Glu Asn Tyr Ala Glu Glu Cys Ala Lys Ala Leu GluLys Gly Leu 65 70 75 80 Pro Asp Pro Val Leu Tyr Leu Ala Glu Lys Phe ThrPro Arg Ser Pro 85 90 95 Cys Gly Leu Tyr Arg Gln Tyr Arg Leu Ala Gly HisTyr Thr Ser Ala 100 105 110 58 22 PRT Homo sapiens 58 Thr Ser Leu ThrSer Pro Cys Pro Leu His Leu Gly Ala Ser Val Leu 1 5 10 15 His Thr HisHis Gly Pro 20 59 19 PRT Homo sapiens 59 Leu Ala Ser Ile Ile Met Ile PheLeu Thr Ala Leu Ala Thr Phe Ile 1 5 10 15 Val Ile Leu 60 20 PRT Homosapiens 60 Leu Phe Trp Leu Leu Arg Val Val Thr Ser Leu Phe Ile Gly AlaAla 1 5 10 15 Ile Leu Ala Val 20 61 22 PRT Homo sapiens 61 Met Leu TrpVal Ala Phe Leu Cys Trp Leu Leu Ala Asn Val Met Leu 1 5 10 15 Ser MetPro Val Leu Val 20 62 17 PRT Homo sapiens 62 Leu Ala Thr Gly Ile Phe GlnLeu Leu Ala Leu Leu Phe Phe Ser Met 1 5 10 15 Ala 63 22 PRT Homo sapiens63 Ala Phe Trp Ile Thr Leu Thr Thr Gly Leu Leu Cys Val Leu Leu Gly 1 510 15 Leu Ala Met Ala Val Ala 20 64 8 PRT Homo sapiens 64 Pro Gly IleArg Gly Lys Thr Arg 1 5 65 6 PRT Homo sapiens 65 Tyr Gly Gly Tyr Met Leu1 5 66 72 PRT Homo sapiens 66 His Arg Met Gln Pro His Arg Leu Lys AlaPhe Phe Asn Gln Ser Val 1 5 10 15 Asp Glu Asp Pro Met Leu Glu Trp SerPro Glu Glu Gly Gly Leu Leu 20 25 30 Ser Pro Arg Tyr Arg Ser Met Ala AspSer Pro Lys Ser Gln Asp Ile 35 40 45 Pro Leu Ser Glu Ala Ser Ser Thr LysAla Tyr Cys Lys Glu Ala His 50 55 60 Pro Lys Asp Pro Asp Cys Ala Leu 6570 67 4928 DNA Mus sp. 67 gtcgacccac gcgtccgccc ggctcccggt gctgccccctctgccccggg ccgcgcccgg 60 gggtcccgca ctgacggccc atggcgccgc ccgccgcccgtctcgcgctg ctctccgccg 120 ctgcgctcac tctggcggcc cggcccgcgc ccggtccccgctccggcccc gagtgcttca 180 cagccaacgg tgcagattac aggggaacac agagctggacagcgctgcaa ggtgggaagc 240 catgtctgtt ctggaacgag actttccagc atccgtacaacacgctgaag taccccaacg 300 gggaaggagg actgggcgag cacaattatt gcagaaatccagatggagac gtgagccctt 360 ggtgctacgt ggccgagcat gaggacggag tctactggaagtactgtgaa attcctgcct 420 gccagatgcc tggaaacctt ggctgctaca aggatcatggaaacccacct cctctcacgg 480 gcaccagtaa aacctctaac aagctcacca tacaaacctgtatcagcttc tgtcggagtc 540 agagattcaa gtttgctggg atggagtcag gctatgcctgcttctgtggg aacaatcctg 600 actactggaa gcacggggag gcggccagca ccgagtgcaatagtgtctgc ttcggggacc 660 acacgcagcc ctgcggtggg gacggcagga ttatcctctttgacactctc gtgggcgcct 720 gcggtgggaa ctactcagcc atggcagccg tggtgtactcccctgacttc cctgacacct 780 acgccactgg cagagtctgc tactggacca tccgggttccaggagcctct cgcatccatt 840 tcaacttcac cctgtttgat atcagggact ctgcagacatggtggagctg ctggacggct 900 acacccaccg cgtcctggtc cggctcagtg ggaggagccgcccgcctctg tctttcaatg 960 tctctctgga ttttgtcatt ttgtatttct tctctgatcgcatcaatcag gcccagggat 1020 ttgctgtgtt gtaccaagcc accaaggagg aaccgccacaggagagacct gctgtcaacc 1080 agaccctggc agaggtgatc accgagcaag ccaacctcagtgtcagcgct gcccactcct 1140 ccaaagtcct ctatgtcatc acccccagcc ccagccaccctccgcagact gcccaggtag 1200 ccattcctgg gcaccgtcag ttggggccaa cagccacagagtggaaggat ggactgtgta 1260 cggcctggcg accctcctca tcctcacagt cacagcagttgtcgcaaaga ttcttctgca 1320 tgtcacattt aaatctcatc gagtccctgc atcaggagaccttagggact gtcgtcagcc 1380 tggggcttct ggagatatct ggaccatttt ctatgaaccttccactacaa tctccatctt 1440 taagaagaag ctcaagggtc agagtcaaca agatgaccgcaatcccctcg tgagtgactg 1500 aagcccacgc ctgcatgaga ggctccgctc caagctcgagtttgctcccc tgagttctcc 1560 tctgatgagt tccctgcctt cccattcacc accatctcttttgggagcac cctgctttag 1620 aggcagccca gcctgggatc ctccatcaca tgtaccagcctggctgctct gctggggatg 1680 gtaagacagg cccaggctga caggacacag ctggacctgactccagaaga ctcttgggtg 1740 gtggggaggt atagtgtagg atgagttttc ttgcttcttctctgttttgt ccacatacag 1800 atcggtttcc cctgtcttta cagtttgcaa tagagccagactgaaagaac tgtcaggttt 1860 tctaggctgg cctggttccc cactaagagt ggcattggcgccctagaggc ccagaggccc 1920 agtgtaggct tggagctttc tctgctgcca actaccatgtgtcatctagt ccgaggggac 1980 tgagagcagg gccacaccag atgtcatctt tctagagggttctttttagt acccactgac 2040 caatggggca agcctgagga ttggtccatc tgtttgtccatggaacagac acagtgaact 2100 tcctggatac tagacttaac tagcctagcc ctcaagtagttgccaatcct gtggaatcag 2160 aattcagcct gtcttcctgt cctcagccca agcctgtagcctagagctgg ggctgtagcc 2220 tagagctggg gctgtagcct agagctgggg ctgtagcacagagctggggc tgtagcctag 2280 agctggggct gtagcacaga gctggggctg tagcctagagctggggctgt agcacagagc 2340 tggggctgta gcacagagct ggggctgtag cctagagctggggctgtagc acagagctgg 2400 ggctgtaact cagcgatcaa gagcttgctt tgtatacatcggaccctagg ttctatccca 2460 gcactatcag aaggtgggag agaaaaagac tgcaccatagcatgcgggca gcatctgtgg 2520 ttcctacgtg aggtgtcatc attttaaaag cagatcaaaactaccgcgag ttttgtcctt 2580 tgtcccttat catgggagca gagtaggagt aagggctctggtcttgctca ttgtccccca 2640 gacagggagg caggaaaagg tcaggcttgg gaactggagatcctcccagg aaaagctgca 2700 agattgagag acccagctgc agttgggaga ggaagggccatccccgactg agaagtcctg 2760 cagtctggaa gtggcctttg tcagcagcag ctgtgccctgaaggtagacc ttggtcactc 2820 tcctgccagc ccttgagcct ctgctctcct gggtaccctcctggaacacc atgctaacct 2880 tccccgagtc tctcagtcac tgccattgag gcctctcctctagctgctgc tccccaggac 2940 tgtctggggc catctgggga tcagggagag gcagcaggagtactgacgag gcagtgacct 3000 gagctgatga gtcaaccaga ggacaccaga gtctacagtgggctggctgc tggctcagct 3060 cctatgggag gcctacaggg gtactaagct agggggtcatcatctcattt gatctgggaa 3120 aggctacagg ctcctggatg tgaagacagg cccactacataagaagacca ctggaaatag 3180 actgacagga gcaggttcca ctctaggctg tccatagcgtttgcaggact cccctgagac 3240 caagtgttga gtcacagagt gccatgtgcg tagtgcataaaggatatggg ttcttaacca 3300 gggaaggctc atagcaggcc aggacatttt ttcagctcagagcactggcc ccaggcttcc 3360 tctaagccac cactcacctg tctcttccta tctcggacacaggaagcaag ccccagtgtg 3420 gtggcagctg cggctcagca ttggtgtccc caggaagggcggtggatgtg cccacgctcc 3480 ttttgctgtg ggcctggcac agcccaacac tgcagggcccaccttctctc ttggggggta 3540 gggacacata aggaaaacta acccacctcc aacaacagcagaggacagtg ggaaggaagg 3600 gctgtaaatc acccaggcca gacctccaga aatgacaggcacagtctgtt agaacctgta 3660 ggcagccagt cacagagggc ctttgtgctg gtaacaccctgcctggagca taggggtaag 3720 ccgagggaga agagcagccc tcagagacat cagctaaaaacataggtgcc ctatgtccct 3780 cccttcctgt cacactgctt acaaagcaga gacagagtaggaaagaggtc ttcatcctct 3840 cccacatcag caaggatagg gctgcggctg cctaaagtgagcaaggagaa cagagctctg 3900 gacttctcta aatgtgggct ctggcttcag actcctcagccaaaagctct tgaagatcaa 3960 agctctggcg ggtacagctg tcctggcctg tgggccagcccatgggatgt gcctgggcca 4020 ggtgccaccc cacggctcac tgtcatccca ggagggaccccacctgatgc tcctcatcat 4080 ccgctggcct gacactatca gagctcgcgc cggctgttgccagggacaga ctgactacac 4140 ttgaccttca agagcactta gaagtggatg gcctccagactctgtcagcc tctgcagggg 4200 ccacacaagt ctcccgagcc aagtccacaa gcctccatggttccctggct cctctcctgt 4260 ggagtgtcct gtttgatgtc tgaggtctgc tttgggtaccgccctgggaa ctgctaacct 4320 ccgattggtc cctttgtgtc tctgtttact gtcctcttctacctccaggt cacttagctc 4380 tggctgctct ggctgggagt ggggggtggg gatgctggctgcacccccac cctggtctgc 4440 caacagaacc tgggggcctc acacgggctc ctgtcttgccaagctggagc tgagcacact 4500 ggcccaggct gagtggggca gagcaaacaa gtggaaggggatctctctcc ttagagggag 4560 gtggccgaag gtgtagatcc agcgagggag ctgccatccccgccaccttc atagcagcaa 4620 gaccttccca tttccaatct caccctccag cagggatatgactttggaca acaaggcttt 4680 atttgtaaat atgctcttaa tatgcaactt tgagaataagatagaaacat catgtatttt 4740 aaaatataaa atgaagtgtg acacactgta tacaatttaatatatatttt taggattttg 4800 ttatttaaga aaatggaatg tgatggtact taacttttacaaaagagaga aaatgttatt 4860 tttactgttt gaagaaaata aatattctca ttgttgtagaaaaaaaaaaa aaaaaaaagg 4920 gcggccgc 4928 68 1410 DNA Mus sp. 68atggcgccgc ccgccgcccg tctcgcgctg ctctccgccg ctgcgctcac tctggcggcc 60cggcccgcgc ccggtccccg ctccggcccc gagtgcttca cagccaacgg tgcagattac 120aggggaacac agagctggac agcgctgcaa ggtgggaagc catgtctgtt ctggaacgag 180actttccagc atccgtacaa cacgctgaag taccccaacg gggaaggagg actgggcgag 240cacaattatt gcagaaatcc agatggagac gtgagccctt ggtgctacgt ggccgagcat 300gaggacggag tctactggaa gtactgtgaa attcctgcct gccagatgcc tggaaacctt 360ggctgctaca aggatcatgg aaacccacct cctctcacgg gcaccagtaa aacctctaac 420aagctcacca tacaaacctg tatcagcttc tgtcggagtc agagattcaa gtttgctggg 480atggagtcag gctatgcctg cttctgtggg aacaatcctg actactggaa gcacggggag 540gcggccagca ccgagtgcaa tagtgtctgc ttcggggacc acacgcagcc ctgcggtggg 600gacggcagga ttatcctctt tgacactctc gtgggcgcct gcggtgggaa ctactcagcc 660atggcagccg tggtgtactc ccctgacttc cctgacacct acgccactgg cagagtctgc 720tactggacca tccgggttcc aggagcctct cgcatccatt tcaacttcac cctgtttgat 780atcagggact ctgcagacat ggtggagctg ctggacggct acacccaccg cgtcctggtc 840cggctcagtg ggaggagccg cccgcctctg tctttcaatg tctctctgga ttttgtcatt 900ttgtatttct tctctgatcg catcaatcag gcccagggat ttgctgtgtt gtaccaagcc 960accaaggagg aaccgccaca ggagagacct gctgtcaacc agaccctggc agaggtgatc 1020accgagcaag ccaacctcag tgtcagcgct gcccactcct ccaaagtcct ctatgtcatc 1080acccccagcc ccagccaccc tccgcagact gcccaggtag ccattcctgg gcaccgtcag 1140ttggggccaa cagccacaga gtggaaggat ggactgtgta cggcctggcg accctcctca 1200tcctcacagt cacagcagtt gtcgcaaaga ttcttctgca tgtcacattt aaatctcatc 1260gagtccctgc atcaggagac cttagggact gtcgtcagcc tggggcttct ggagatatct 1320ggaccatttt ctatgaacct tccactacaa tctccatctt taagaagaag ctcaagggtc 1380agagtcaaca agatgaccgc aatcccctcg 1410 69 470 PRT Mus sp. 69 Met Ala ProPro Ala Ala Arg Leu Ala Leu Leu Ser Ala Ala Ala Leu 1 5 10 15 Thr LeuAla Ala Arg Pro Ala Pro Gly Pro Arg Ser Gly Pro Glu Cys 20 25 30 Phe ThrAla Asn Gly Ala Asp Tyr Arg Gly Thr Gln Ser Trp Thr Ala 35 40 45 Leu GlnGly Gly Lys Pro Cys Leu Phe Trp Asn Glu Thr Phe Gln His 50 55 60 Pro TyrAsn Thr Leu Lys Tyr Pro Asn Gly Glu Gly Gly Leu Gly Glu 65 70 75 80 HisAsn Tyr Cys Arg Asn Pro Asp Gly Asp Val Ser Pro Trp Cys Tyr 85 90 95 ValAla Glu His Glu Asp Gly Val Tyr Trp Lys Tyr Cys Glu Ile Pro 100 105 110Ala Cys Gln Met Pro Gly Asn Leu Gly Cys Tyr Lys Asp His Gly Asn 115 120125 Pro Pro Pro Leu Thr Gly Thr Ser Lys Thr Ser Asn Lys Leu Thr Ile 130135 140 Gln Thr Cys Ile Ser Phe Cys Arg Ser Gln Arg Phe Lys Phe Ala Gly145 150 155 160 Met Glu Ser Gly Tyr Ala Cys Phe Cys Gly Asn Asn Pro AspTyr Trp 165 170 175 Lys His Gly Glu Ala Ala Ser Thr Glu Cys Asn Ser ValCys Phe Gly 180 185 190 Asp His Thr Gln Pro Cys Gly Gly Asp Gly Arg IleIle Leu Phe Asp 195 200 205 Thr Leu Val Gly Ala Cys Gly Gly Asn Tyr SerAla Met Ala Ala Val 210 215 220 Val Tyr Ser Pro Asp Phe Pro Asp Thr TyrAla Thr Gly Arg Val Cys 225 230 235 240 Tyr Trp Thr Ile Arg Val Pro GlyAla Ser Arg Ile His Phe Asn Phe 245 250 255 Thr Leu Phe Asp Ile Arg AspSer Ala Asp Met Val Glu Leu Leu Asp 260 265 270 Gly Tyr Thr His Arg ValLeu Val Arg Leu Ser Gly Arg Ser Arg Pro 275 280 285 Pro Leu Ser Phe AsnVal Ser Leu Asp Phe Val Ile Leu Tyr Phe Phe 290 295 300 Ser Asp Arg IleAsn Gln Ala Gln Gly Phe Ala Val Leu Tyr Gln Ala 305 310 315 320 Thr LysGlu Glu Pro Pro Gln Glu Arg Pro Ala Val Asn Gln Thr Leu 325 330 335 AlaGlu Val Ile Thr Glu Gln Ala Asn Leu Ser Val Ser Ala Ala His 340 345 350Ser Ser Lys Val Leu Tyr Val Ile Thr Pro Ser Pro Ser His Pro Pro 355 360365 Gln Thr Ala Gln Val Ala Ile Pro Gly His Arg Gln Leu Gly Pro Thr 370375 380 Ala Thr Glu Trp Lys Asp Gly Leu Cys Thr Ala Trp Arg Pro Ser Ser385 390 395 400 Ser Ser Gln Ser Gln Gln Leu Ser Gln Arg Phe Phe Cys MetSer His 405 410 415 Leu Asn Leu Ile Glu Ser Leu His Gln Glu Thr Leu GlyThr Val Val 420 425 430 Ser Leu Gly Leu Leu Glu Ile Ser Gly Pro Phe SerMet Asn Leu Pro 435 440 445 Leu Gln Ser Pro Ser Leu Arg Arg Ser Ser ArgVal Arg Val Asn Lys 450 455 460 Met Thr Ala Ile Pro Ser 465 470 70 760PRT Mus sp. 70 Met Ala Leu Pro Ser Leu Gly Gln Asp Ser Trp Ser Leu LeuArg Val 1 5 10 15 Phe Phe Phe Gln Leu Phe Leu Leu Pro Ser Leu Pro ProAla Ser Gly 20 25 30 Thr Gly Gly Gln Gly Pro Met Pro Arg Val Lys Tyr HisAla Gly Asp 35 40 45 Gly His Arg Ala Leu Ser Phe Phe Gln Gln Lys Gly LeuArg Asp Phe 50 55 60 Asp Thr Leu Leu Leu Ser Asp Asp Gly Asn Thr Leu TyrVal Gly Ala 65 70 75 80 Arg Glu Thr Val Leu Ala Leu Asn Ile Gln Asn ProGly Ile Pro Arg 85 90 95 Leu Lys Asn Met Ile Pro Trp Pro Ala Ser Glu ArgLys Lys Thr Glu 100 105 110 Cys Ala Phe Lys Lys Lys Ser Asn Glu Thr GlnCys Phe Asn Phe Ile 115 120 125 Arg Val Leu Val Ser Tyr Asn Ala Thr HisLeu Tyr Ala Cys Gly Thr 130 135 140 Phe Ala Phe Ser Pro Ala Cys Thr PheIle Glu Leu Gln Asp Ser Leu 145 150 155 160 Leu Leu Pro Ile Leu Ile AspLys Val Met Asp Gly Lys Gly Gln Ser 165 170 175 Pro Leu Thr Leu Phe ThrSer Thr Gln Ala Val Leu Val Asp Gly Met 180 185 190 Leu Tyr Ser Gly ThrMet Asn Asn Phe Leu Gly Ser Glu Pro Ile Leu 195 200 205 Met Arg Thr LeuGly Ser His Pro Val Leu Lys Thr Asp Ile Phe Leu 210 215 220 Arg Trp LeuHis Ala Asp Ala Ser Phe Val Ala Ala Ile Pro Ser Thr 225 230 235 240 GlnVal Val Tyr Phe Phe Phe Glu Glu Thr Ala Ser Glu Phe Asp Phe 245 250 255Phe Glu Glu Leu Tyr Ile Ser Arg Val Ala Gln Val Cys Lys Asn Asp 260 265270 Val Gly Gly Glu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe Leu Lys 275280 285 Ala Gln Leu Leu Cys Ala Gln Pro Gly Gln Leu Pro Phe Asn Ile Ile290 295 300 Arg His Ala Val Leu Leu Pro Ala Asp Ser Pro Ser Val Ser ArgIle 305 310 315 320 Tyr Ala Val Phe Thr Ser Gln Trp Gln Val Gly Gly ThrArg Ser Ser 325 330 335 Ala Val Cys Ala Phe Ser Leu Thr Asp Ile Glu ArgVal Phe Lys Gly 340 345 350 Lys Tyr Lys Glu Leu Asn Lys Glu Thr Ser ArgTrp Thr Thr Tyr Arg 355 360 365 Gly Ser Glu Val Ser Pro Arg Pro Gly SerCys Ser Met Gly Pro Ser 370 375 380 Ser Asp Lys Ala Leu Thr Phe Met LysAsp His Phe Leu Met Asp Glu 385 390 395 400 His Val Val Gly Thr Pro LeuLeu Val Lys Ser Gly Val Glu Tyr Thr 405 410 415 Arg Leu Ala Val Glu SerAla Arg Gly Leu Asp Gly Ser Ser His Val 420 425 430 Val Met Tyr Leu GlyThr Ser Thr Gly Pro Leu His Lys Ala Val Val 435 440 445 Pro Gln Asp SerSer Ala Tyr Leu Val Glu Glu Ile Gln Leu Ser Pro 450 455 460 Asp Ser GluPro Val Arg Asn Leu Gln Leu Ala Pro Ala Gln Gly Ala 465 470 475 480 ValPhe Ala Gly Phe Ser Gly Gly Ile Trp Arg Val Pro Arg Ala Asn 485 490 495Cys Ser Val Tyr Glu Ser Cys Val Asp Cys Val Leu Ala Arg Asp Pro 500 505510 His Cys Ala Trp Asp Pro Glu Ser Arg Leu Cys Ser Leu Leu Ser Gly 515520 525 Ser Thr Lys Pro Trp Lys Gln Asp Met Glu Arg Gly Asn Pro Glu Trp530 535 540 Val Cys Thr Arg Gly Pro Met Ala Arg Ser Pro Arg Arg Gln SerPro 545 550 555 560 Pro Gln Leu Ile Lys Glu Val Leu Thr Val Pro Asn SerIle Leu Glu 565 570 575 Leu Arg Cys Pro His Leu Ser Ala Leu Ala Ser TyrHis Trp Ser His 580 585 590 Gly Arg Ala Lys Ile Ser Glu Ala Ser Ala ThrVal Tyr Asn Gly Ser 595 600 605 Leu Leu Leu Leu Pro Gln Asp Gly Val GlyGly Leu Tyr Gln Cys Val 610 615 620 Ala Thr Glu Asn Gly Tyr Ser Tyr ProVal Val Ser Tyr Trp Val Asp 625 630 635 640 Ser Gln Asp Gln Pro Leu AlaLeu Asp Pro Glu Leu Ala Gly Val Pro 645 650 655 Arg Glu Arg Val Gln ValPro Leu Thr Arg Val Gly Gly Gly Ala Ser 660 665 670 Met Ala Ala Gln ArgSer Tyr Trp Pro His Phe Leu Ile Val Thr Val 675 680 685 Leu Leu Ala IleVal Leu Leu Gly Val Leu Thr Leu Leu Leu Ala Ser 690 695 700 Pro Leu GlyAla Leu Arg Ala Arg Gly Lys Val Gln Gly Cys Gly Met 705 710 715 720 LeuPro Pro Arg Glu Lys Ala Pro Leu Ser Arg Asp Gln His Leu Gln 725 730 735Pro Ser Lys Asp His Arg Thr Ser Ala Ser Asp Val Asp Ala Asp Asn 740 745750 Asn His Leu Gly Ala Glu Val Ala 755 760 71 3046 DNA Mus sp. 71ctcggacgcc tgggttaggg gtctgtactg ctggggaacc atctggtgac catctcaggc 60tgaccatggc cctaccatcc ctgggccagg actcatggag tctcctgcgt gtttttttct 120tccaactctt cctgctgcca tcactgccac ctgcttctgg gactggtggt caggggccca 180tgcccagagt caaataccat gctggagacg ggcacagggc cctcagcttc ttccaacaaa 240aaggcctccg agactttgac acgctgctcc tgagtgacga tggcaacact ctctatgtgg 300gggctcgaga gaccgtcctg gccttgaata tccagaaccc aggaatccca aggctaaaga 360acatgatacc ctggccagcc agtgagagaa aaaagaccga atgtgccttt aagaagaaga 420gcaatgagac acagtgtttc aacttcattc gagtcctggt ctcttacaat gctactcacc 480tctatgcctg tgggaccttt gccttcagcc ctgcctgtac cttcattgaa ctccaagatt 540ccctcctgtt gcccatcttg atagacaagg tcatggacgg gaagggccaa agccctttga 600ccctgttcac aagcacacaa gctgtcttgg tcgatgggat gctttattcc ggcaccatga 660acaacttcct gggcagcgag cccatcctga tgcggacact gggatcccat cctgttctca 720agactgacat cttcttacgc tggctgcacg cggatgcctc cttcgtggca gccattccat 780ccacccaggt cgtctatttc ttctttgagg agacagccag cgagtttgac ttctttgaag 840agctgtatat atccagggtg gctcaagtct gcaagaacga cgtgggcggt gaaaagctgc 900tgcagaagaa gtggaccacc ttcctcaaag cccagttgct ctgcgctcag ccagggcagc 960tgccattcaa catcatccgc cacgcggtcc tgctgcccgc cgattctccc tctgtttccc 1020gcatctacgc agtctttacc tcccagtggc aggttggcgg gaccaggagc tcagcagtct 1080gtgccttctc tctcacggac attgagcgag tctttaaagg gaagtacaag gagctgaaca 1140aggagacctc ccgctggacc acttaccggg gctcagaggt cagcccgagg ccaggcagtt 1200gctccatggg cccctcctct gacaaagcct tgaccttcat gaaggaccat tttctgatgg 1260atgagcacgt ggtaggaaca cccctgctgg tgaagtctgg tgtggagtac acacggcttg 1320ctgtggagtc agctcggggc cttgatggga gcagccatgt ggtcatgtat ctgggtacct 1380ccacgggtcc cctgcacaag gctgtggtgc ctcaggacag cagtgcttat ctcgtggagg 1440agattcagct gagccctgac tctgagcctg ttcgaaacct gcagctggcc cccgcccagg 1500gtgcagtgtt tgcaggcttc tctggaggca tctggagagt tcccagggcc aattgcagtg 1560tctacgagag ctgtgtggac tgtgtgcttg ccagggaccc tcactgtgcc tgggaccctg 1620aatcaagact ctgcagcctt ctgtctggct ctaccaagcc ttggaagcag gacatggaac 1680gcggcaaccc ggagtgggta tgcacccgtg gccccatggc caggagcccc cggcgtcaga 1740gcccccctca actaattaaa gaagtcctga cagtccccaa ctccatcctg gagctgcgct 1800gcccccacct gtcagcactg gcctcttacc actggagtca tggccgagcc aaaatctcag 1860aagcctctgc taccgtctac aatggctccc tcttgctgct gccgcaggat ggtgtcgggg 1920gcctctacca gtgtgtggcg actgagaacg gctactcata ccctgtggtc tcctattggg 1980tagacagcca ggaccagccc ctggcgctgg accctgagct ggcgggcgtt ccccgtgagc 2040gtgtgcaggt cccgctgacc agggtcggag gcggagcttc catggctgcc cagcggtcct 2100actggcccca ttttctcatc gttaccgtcc tcctggccat cgtgctcctg ggagtgctca 2160ctctcctcct cgcttcccca ctgggggcgc tgcgggctcg gggtaaggtt cagggctgtg 2220ggatgctgcc ccccagggaa aaggctccac tgagcaggga ccagcacctc cagccctcca 2280aggaccacag gacctctgcc agtgacgtag atgccgacaa caaccatctg ggcgccgaag 2340tggcttaaac agggacacag atccgcagct gagcagagca agccactggc cttgttggct 2400atgccaggca cagtgccact ctgaccaggg taggaggctc tcctgctaac gtgtgtcacc 2460tacagcaccc agtaggtcct cccctgtggg actctcttct gcaagcacat tgggctgtct 2520ccatacctgt acttgtgctg tgacaggaag agccagacag gtttctttga ttttgattga 2580cccaagagcc ctgcctgtaa caaacgtgct ccaggagacc atgaaaggtg tggctgtctg 2640ggattctgtg gtgacaaacc taagcatccg agcaagctgg ggctattcct gcaaactcca 2700tcctgaacgc tgtcactcta gaagcagctg ctgctttgaa caccagccca ccctccttcc 2760caagagtctc tatggagttg gccccttgtg tttcctttac cagtcgggcc atactgtttg 2820ggaagtcatc tctgaagtct aaccaccttc cttcttggtt cagtttggac agattgttat 2880tattgtctct gccctggcta gaatgggggc ataatctgag ccttgttccc ttgtccagtg 2940tggctgaccc ttgacctctt ccttcctcct ccctttgttt tgggattcag aaaactgctt 3000gtcacagaca atttattttt tattaaaaaa gatataagct ttaaag 3046 72 2915 DNA Mussp. 72 gtcgacccac gcgtccggcc gcgcgtcctt ctgccggctt cagctcgtat ccccggagtc60 cacccgcccg tcccggggtg cggactggcc ctgagctggc cgtacagccc ggcttcggac 120ggtcctcgct ggagccatgg gccgccggct cggcagggtg gcggcgctgc tgctcgggct 180gctagtggag tgcactgagg ccaaaaaaca ttgctggtat tttgaaggac tctatcccac 240atactatata tgccgttcct atgaagactg ctgtggctcc aggtgctgtg tgagggccct 300ttccatacag aggctgtggt atttttggtt cctgctgatg atgggtgtgc tgttctgctg 360tggtgccggt ttcttcattc gccggcgcat gtatccgcca ccactcattg aggagcccac 420attcaatgtg tcctatacca ggcagccacc aaatcctgct ccaggagcac agcaaatggg 480accgccatat tacaccgacc ctggaggacc cgggatgaat cctgttggca ataccatggc 540tatggctttc caggtccagc ccaattcacc tcacggaggc acaacttacc caccccctcc 600ttcctactgc aacacgcctc caccccccta tgaacaggtg gtgaaggaca agtagcaaga 660tgctacatca aaggcaaaga ggatggacag gcccttttgt ttaccttccc atcctcaccg 720atacttgctg atagggtggt ccaagggaaa acttggatat tctcaaagca agcccagctc 780tctttcaagt cttttgtgga ggacatttga atccacactg tctcctctgt tgcttctgtt 840tctgatgtag tctgtgctct ctgagagagt gtggcaacag tccctgaggg ttgatattcc 900tagggtgtcc agggtagatc ctcgggagag aggctaaggg gaaaggaagg catagcctgt 960gtgttagggg gcagataaag tggtcaggct gagataagac tcacatgatg cagtagttgg 1020cagtgaactt cgaagagaca ctatccacca tcccagccca ttctcctaat agaagctgtg 1080gggctgtgtt gttgatgctc tttggtctcc actcacattt tgaaaatagg ctttcctctg 1140caggaatagg aaagacccaa gtacatattt gcttccactt aaaaatgagg gtcagaacca 1200ggcctcagtt ggacatctat agttaaataa aggccattag agaggggaaa tctttaagtt 1260aggggaaatt ctctaaatgg agacattgcg ttttatgaat catcgtctgg cttttctttt 1320agtgcatgta ttgaagtgag ggtgtccttt gagatcagat ggggagagtg aactctgcgg 1380ggggtggggt gtctctactc agagggctcc aacacccttt tcttaggtag ttctggtgat 1440gggttttatg ggcactatag agctgagggg cacattaggc cgggtagtta cattgaccct 1500tggagaggaa gaggacagcc aaagaaactc agcaaagcaa gaccagcatt gctgagttag 1560agctagggtt gtatgtgatc ccaacagaga tgtgctggcc tcagaagagg ggacgtttgt 1620ggatagagcc gtgaaaacct acttagttgc acagatgaca taatcaaaag tagagaaaga 1680agtgtagtta gagatgccat ttcccaggtg agaatcagag ctcatccata gatttacaag 1740tagtggctgg agttaacagt atggagttct tttcccttgc gtagttagtc acgttgatgt 1800gtatttaaac ccaggttgag accttgtgta ctaagagcaa ggaagtatag ctaagatgtc 1860tagattattt atatgtagta tggtggggag tggggctgca aggaaggggg ctgacattgt 1920aaatgagaaa atcagagcca tttgataaac tgttacttgt tggatcaggc atccaaaagt 1980gtctcttgag tggacattga gtattcttta ccacctacaa gaccaggagg catggtgtca 2040ttctccattg gggtatttat atgaggtaga ggttcaggaa tcgacagtag ctgtgtgggc 2100ttagtttaag gactgaaagc atagggactg gtagacagtt tcataggaaa ctgcggggaa 2160ggaatggata cctttaaaga cagtttgtgg atgcagatgc tgccacccat cattgagcac 2220ccttgtgtct ctggcttcct gtcactggat ccagtacccc tccatgcttg ggtccttgtt 2280ttacataaga caacaaagca caatgtctgc tgtttacaat caagacgact acatggtcca 2340aacatttctt ctctcttcta tcacttgtgg ctttaacttc catttcctcc gttccttttt 2400aaaatcaaga agcacagtca gagctgcccc tgggattgca tcagggaacg gctgatcaag 2460gcattcagtg tccatgacta aatcttatct ttttgatagc aaatcctttt aagaaactga 2520acaattgcta aggctcagca attttatact ccaatgtctg tgtaaggtaa attttgtttg 2580ccattgagcc cacattggaa ttccttctga cgtcaacact gacaatgcct atggaaattg 2640cacttctggg tatatgtccc agcatccttg ttttcttatg tttggtgagt aaggctcacc 2700ccttccagca gctctacttc tgtgtgctga ggtcctgtag agccggggct tgggcacaga 2760catgaggcag acttgtgcat gctctttctt ggcaacactt ggctcatatt tcttgttctc 2820ttttgataga gtcctgtttc ctatgtattt aaaaaataat aaaagtgaat ttagtcaaaa 2880aaaaaaaaaa aaaaaaaaaa aaaaagggcg gccgc 2915 73 516 DNA Mus sp. 73atgggccgcc ggctcggcag ggtggcggcg ctgctgctcg ggctgctagt ggagtgcact 60gaggccaaaa aacattgctg gtattttgaa ggactctatc ccacatacta tatatgccgt 120tcctatgaag actgctgtgg ctccaggtgc tgtgtgaggg ccctttccat acagaggctg 180tggtattttt ggttcctgct gatgatgggt gtgctgttct gctgtggtgc cggtttcttc 240attcgccggc gcatgtatcc gccaccactc attgaggagc ccacattcaa tgtgtcctat 300accaggcagc caccaaatcc tgctccagga gcacagcaaa tgggaccgcc atattacacc 360gaccctggag gacccgggat gaatcctgtt ggcaatacca tggctatggc tttccaggtc 420cagcccaatt cacctcacgg aggcacaact tacccacccc ctccttccta ctgcaacacg 480cctccacccc cctatgaaca ggtggtgaag gacaag 516 74 172 PRT Mus sp. 74 MetGly Arg Arg Leu Gly Arg Val Ala Ala Leu Leu Leu Gly Leu Leu 1 5 10 15Val Glu Cys Thr Glu Ala Lys Lys His Cys Trp Tyr Phe Glu Gly Leu 20 25 30Tyr Pro Thr Tyr Tyr Ile Cys Arg Ser Tyr Glu Asp Cys Cys Gly Ser 35 40 45Arg Cys Cys Val Arg Ala Leu Ser Ile Gln Arg Leu Trp Tyr Phe Trp 50 55 60Phe Leu Leu Met Met Gly Val Leu Phe Cys Cys Gly Ala Gly Phe Phe 65 70 7580 Ile Arg Arg Arg Met Tyr Pro Pro Pro Leu Ile Glu Glu Pro Thr Phe 85 9095 Asn Val Ser Tyr Thr Arg Gln Pro Pro Asn Pro Ala Pro Gly Ala Gln 100105 110 Gln Met Gly Pro Pro Tyr Tyr Thr Asp Pro Gly Gly Pro Gly Met Asn115 120 125 Pro Val Gly Asn Thr Met Ala Met Ala Phe Gln Val Gln Pro AsnSer 130 135 140 Pro His Gly Gly Thr Thr Tyr Pro Pro Pro Pro Ser Tyr CysAsn Thr 145 150 155 160 Pro Pro Pro Pro Tyr Glu Gln Val Val Lys Asp Lys165 170 75 398 PRT Homo sapiens 75 Met Trp Leu Leu Leu Thr Met Ala SerLeu Ile Ser Val Leu Gly Thr 1 5 10 15 Thr His Gly Leu Phe Gly Lys LeuHis Pro Gly Ser Pro Glu Val Thr 20 25 30 Met Asn Ile Ser Gln Met Ile ThrTyr Trp Gly Tyr Pro Asn Glu Glu 35 40 45 Tyr Glu Val Val Thr Glu Asp GlyTyr Ile Leu Glu Val Asn Arg Ile 50 55 60 Pro Tyr Gly Lys Lys Asn Ser GlyAsn Thr Gly Gln Arg Pro Val Val 65 70 75 80 Phe Leu Gln His Gly Leu LeuAla Ser Ala Thr Asn Trp Ile Ser Asn 85 90 95 Leu Pro Asn Asn Ser Leu AlaPhe Ile Leu Ala Asp Ala Gly Tyr Asp 100 105 110 Val Trp Leu Gly Asn SerArg Gly Asn Thr Trp Ala Arg Arg Asn Leu 115 120 125 Tyr Tyr Ser Pro AspSer Val Glu Phe Trp Ala Phe Ser Phe Asp Glu 130 135 140 Met Ala Lys TyrAsp Leu Pro Ala Thr Ile Asp Phe Ile Val Lys Lys 145 150 155 160 Thr GlyGln Lys Gln Leu His Tyr Val Gly His Ser Gln Gly Thr Thr 165 170 175 IleGly Phe Ile Ala Phe Ser Thr Asn Pro Ser Leu Ala Lys Arg Ile 180 185 190Lys Thr Phe Tyr Ala Leu Ala Pro Val Ala Thr Val Lys Tyr Thr Lys 195 200205 Ser Leu Ile Asn Lys Leu Arg Phe Val Pro Gln Ser Leu Phe Lys Phe 210215 220 Ile Phe Gly Asp Lys Ile Phe Tyr Pro His Asn Phe Phe Asp Gln Phe225 230 235 240 Leu Ala Thr Glu Val Cys Ser Arg Glu Met Leu Asn Leu LeuCys Ser 245 250 255 Asn Ala Leu Phe Ile Ile Cys Gly Phe Asp Ser Lys AsnPhe Asn Thr 260 265 270 Ser Arg Leu Asp Val Tyr Leu Ser His Asn Pro AlaGly Thr Ser Val 275 280 285 Gln Asn Met Phe His Trp Thr Gln Ala Val LysSer Gly Lys Phe Gln 290 295 300 Ala Tyr Asp Trp Gly Ser Pro Val Gln AsnArg Met His Tyr Asp Gln 305 310 315 320 Ser Gln Pro Pro Tyr Tyr Asn ValThr Ala Met Asn Val Pro Ile Ala 325 330 335 Val Trp Asn Gly Gly Lys AspLeu Leu Ala Asp Pro Gln Asp Val Gly 340 345 350 Leu Leu Leu Pro Lys LeuPro Asn Leu Ile Tyr His Lys Glu Ile Pro 355 360 365 Phe Tyr Asn His LeuAsp Phe Ile Trp Ala Met Asp Ala Pro Gln Glu 370 375 380 Val Tyr Asn AspIle Val Ser Met Ile Ser Glu Asp Lys Lys 385 390 395 76 760 PRT Mus sp.76 Met Ala Leu Pro Ser Leu Gly Gln Asp Ser Trp Ser Leu Leu Arg Val 1 510 15 Phe Phe Phe Gln Leu Phe Leu Leu Pro Ser Leu Pro Pro Ala Ser Gly 2025 30 Thr Gly Gly Gln Gly Pro Met Pro Arg Val Lys Tyr His Ala Gly Asp 3540 45 Gly His Arg Ala Leu Ser Phe Phe Gln Gln Lys Gly Leu Arg Asp Phe 5055 60 Asp Thr Leu Leu Leu Ser Asp Asp Gly Asn Thr Leu Tyr Val Gly Ala 6570 75 80 Arg Glu Thr Val Leu Ala Leu Asn Ile Gln Asn Pro Gly Ile Pro Arg85 90 95 Leu Lys Asn Met Ile Pro Trp Pro Ala Ser Glu Arg Lys Lys Thr Glu100 105 110 Cys Ala Phe Lys Lys Lys Ser Asn Glu Thr Gln Cys Phe Asn PheIle 115 120 125 Arg Val Leu Val Ser Tyr Asn Ala Thr His Leu Tyr Ala CysGly Thr 130 135 140 Phe Ala Phe Ser Pro Ala Cys Thr Phe Ile Glu Leu GlnAsp Ser Leu 145 150 155 160 Leu Leu Pro Ile Leu Ile Asp Lys Val Met AspGly Lys Gly Gln Ser 165 170 175 Pro Leu Thr Leu Phe Thr Ser Thr Gln AlaVal Leu Val Asp Gly Met 180 185 190 Leu Tyr Ser Gly Thr Met Asn Asn PheLeu Gly Ser Glu Pro Ile Leu 195 200 205 Met Arg Thr Leu Gly Ser His ProVal Leu Lys Thr Asp Ile Phe Leu 210 215 220 Arg Trp Leu His Ala Asp AlaSer Phe Val Ala Ala Ile Pro Ser Thr 225 230 235 240 Gln Val Val Tyr PhePhe Phe Glu Glu Thr Ala Ser Glu Phe Asp Phe 245 250 255 Phe Glu Glu LeuTyr Ile Ser Arg Val Ala Gln Val Cys Lys Asn Asp 260 265 270 Val Gly GlyGlu Lys Leu Leu Gln Lys Lys Trp Thr Thr Phe Leu Lys 275 280 285 Ala GlnLeu Leu Cys Ala Gln Pro Gly Gln Leu Pro Phe Asn Ile Ile 290 295 300 ArgHis Ala Val Leu Leu Pro Ala Asp Ser Pro Ser Val Ser Arg Ile 305 310 315320 Tyr Ala Val Phe Thr Ser Gln Trp Gln Val Gly Gly Thr Arg Ser Ser 325330 335 Ala Val Cys Ala Phe Ser Leu Thr Asp Ile Glu Arg Val Phe Lys Gly340 345 350 Lys Tyr Lys Glu Leu Asn Lys Glu Thr Ser Arg Trp Thr Thr TyrArg 355 360 365 Gly Ser Glu Val Ser Pro Arg Pro Gly Ser Cys Ser Met GlyPro Ser 370 375 380 Ser Asp Lys Ala Leu Thr Phe Met Lys Asp His Phe LeuMet Asp Glu 385 390 395 400 His Val Val Gly Thr Pro Leu Leu Val Lys SerGly Val Glu Tyr Thr 405 410 415 Arg Leu Ala Val Glu Ser Ala Arg Gly LeuAsp Gly Ser Ser His Val 420 425 430 Val Met Tyr Leu Gly Thr Ser Thr GlyPro Leu His Lys Ala Val Val 435 440 445 Pro Gln Asp Ser Ser Ala Tyr LeuVal Glu Glu Ile Gln Leu Ser Pro 450 455 460 Asp Ser Glu Pro Val Arg AsnLeu Gln Leu Ala Pro Ala Gln Gly Ala 465 470 475 480 Val Phe Ala Gly PheSer Gly Gly Ile Trp Arg Val Pro Arg Ala Asn 485 490 495 Cys Ser Val TyrGlu Ser Cys Val Asp Cys Val Leu Ala Arg Asp Pro 500 505 510 His Cys AlaTrp Asp Pro Glu Ser Arg Leu Cys Ser Leu Leu Ser Gly 515 520 525 Ser ThrLys Pro Trp Lys Gln Asp Met Glu Arg Gly Asn Pro Glu Trp 530 535 540 ValCys Thr Arg Gly Pro Met Ala Arg Ser Pro Arg Arg Gln Ser Pro 545 550 555560 Pro Gln Leu Ile Lys Glu Val Leu Thr Val Pro Asn Ser Ile Leu Glu 565570 575 Leu Arg Cys Pro His Leu Ser Ala Leu Ala Ser Tyr His Trp Ser His580 585 590 Gly Arg Ala Lys Ile Ser Glu Ala Ser Ala Thr Val Tyr Asn GlySer 595 600 605 Leu Leu Leu Leu Pro Gln Asp Gly Val Gly Gly Leu Tyr GlnCys Val 610 615 620 Ala Thr Glu Asn Gly Tyr Ser Tyr Pro Val Val Ser TyrTrp Val Asp 625 630 635 640 Ser Gln Asp Gln Pro Leu Ala Leu Asp Pro GluLeu Ala Gly Val Pro 645 650 655 Arg Glu Arg Val Gln Val Pro Leu Thr ArgVal Gly Gly Gly Ala Ser 660 665 670 Met Ala Ala Gln Arg Ser Tyr Trp ProHis Phe Leu Ile Val Thr Val 675 680 685 Leu Leu Ala Ile Val Leu Leu GlyVal Leu Thr Leu Leu Leu Ala Ser 690 695 700 Pro Leu Gly Ala Leu Arg AlaArg Gly Lys Val Gln Gly Cys Gly Met 705 710 715 720 Leu Pro Pro Arg GluLys Ala Pro Leu Ser Arg Asp Gln His Leu Gln 725 730 735 Pro Ser Lys AspHis Arg Thr Ser Ala Ser Asp Val Asp Ala Asp Asn 740 745 750 Asn His LeuGly Ala Glu Val Ala 755 760 77 3046 DNA Mus sp. 77 ctcggacgcc tgggttaggggtctgtactg ctggggaacc atctggtgac catctcaggc 60 tgaccatggc cctaccatccctgggccagg actcatggag tctcctgcgt gtttttttct 120 tccaactctt cctgctgccatcactgccac ctgcttctgg gactggtggt caggggccca 180 tgcccagagt caaataccatgctggagacg ggcacagggc cctcagcttc ttccaacaaa 240 aaggcctccg agactttgacacgctgctcc tgagtgacga tggcaacact ctctatgtgg 300 gggctcgaga gaccgtcctggccttgaata tccagaaccc aggaatccca aggctaaaga 360 acatgatacc ctggccagccagtgagagaa aaaagaccga atgtgccttt aagaagaaga 420 gcaatgagac acagtgtttcaacttcattc gagtcctggt ctcttacaat gctactcacc 480 tctatgcctg tgggacctttgccttcagcc ctgcctgtac cttcattgaa ctccaagatt 540 ccctcctgtt gcccatcttgatagacaagg tcatggacgg gaagggccaa agccctttga 600 ccctgttcac aagcacacaagctgtcttgg tcgatgggat gctttattcc ggcaccatga 660 acaacttcct gggcagcgagcccatcctga tgcggacact gggatcccat cctgttctca 720 agactgacat cttcttacgctggctgcacg cggatgcctc cttcgtggca gccattccat 780 ccacccaggt cgtctatttcttctttgagg agacagccag cgagtttgac ttctttgaag 840 agctgtatat atccagggtggctcaagtct gcaagaacga cgtgggcggt gaaaagctgc 900 tgcagaagaa gtggaccaccttcctcaaag cccagttgct ctgcgctcag ccagggcagc 960 tgccattcaa catcatccgccacgcggtcc tgctgcccgc cgattctccc tctgtttccc 1020 gcatctacgc agtctttacctcccagtggc aggttggcgg gaccaggagc tcagcagtct 1080 gtgccttctc tctcacggacattgagcgag tctttaaagg gaagtacaag gagctgaaca 1140 aggagacctc ccgctggaccacttaccggg gctcagaggt cagcccgagg ccaggcagtt 1200 gctccatggg cccctcctctgacaaagcct tgaccttcat gaaggaccat tttctgatgg 1260 atgagcacgt ggtaggaacacccctgctgg tgaagtctgg tgtggagtac acacggcttg 1320 ctgtggagtc agctcggggccttgatggga gcagccatgt ggtcatgtat ctgggtacct 1380 ccacgggtcc cctgcacaaggctgtggtgc ctcaggacag cagtgcttat ctcgtggagg 1440 agattcagct gagccctgactctgagcctg ttcgaaacct gcagctggcc cccgcccagg 1500 gtgcagtgtt tgcaggcttctctggaggca tctggagagt tcccagggcc aattgcagtg 1560 tctacgagag ctgtgtggactgtgtgcttg ccagggaccc tcactgtgcc tgggaccctg 1620 aatcaagact ctgcagccttctgtctggct ctaccaagcc ttggaagcag gacatggaac 1680 gcggcaaccc ggagtgggtatgcacccgtg gccccatggc caggagcccc cggcgtcaga 1740 gcccccctca actaattaaagaagtcctga cagtccccaa ctccatcctg gagctgcgct 1800 gcccccacct gtcagcactggcctcttacc actggagtca tggccgagcc aaaatctcag 1860 aagcctctgc taccgtctacaatggctccc tcttgctgct gccgcaggat ggtgtcgggg 1920 gcctctacca gtgtgtggcgactgagaacg gctactcata ccctgtggtc tcctattggg 1980 tagacagcca ggaccagcccctggcgctgg accctgagct ggcgggcgtt ccccgtgagc 2040 gtgtgcaggt cccgctgaccagggtcggag gcggagcttc catggctgcc cagcggtcct 2100 actggcccca ttttctcatcgttaccgtcc tcctggccat cgtgctcctg ggagtgctca 2160 ctctcctcct cgcttccccactgggggcgc tgcgggctcg gggtaaggtt cagggctgtg 2220 ggatgctgcc ccccagggaaaaggctccac tgagcaggga ccagcacctc cagccctcca 2280 aggaccacag gacctctgccagtgacgtag atgccgacaa caaccatctg ggcgccgaag 2340 tggcttaaac agggacacagatccgcagct gagcagagca agccactggc cttgttggct 2400 atgccaggca cagtgccactctgaccaggg taggaggctc tcctgctaac gtgtgtcacc 2460 tacagcaccc agtaggtcctcccctgtggg actctcttct gcaagcacat tgggctgtct 2520 ccatacctgt acttgtgctgtgacaggaag agccagacag gtttctttga ttttgattga 2580 cccaagagcc ctgcctgtaacaaacgtgct ccaggagacc atgaaaggtg tggctgtctg 2640 ggattctgtg gtgacaaacctaagcatccg agcaagctgg ggctattcct gcaaactcca 2700 tcctgaacgc tgtcactctagaagcagctg ctgctttgaa caccagccca ccctccttcc 2760 caagagtctc tatggagttggccccttgtg tttcctttac cagtcgggcc atactgtttg 2820 ggaagtcatc tctgaagtctaaccaccttc cttcttggtt cagtttggac agattgttat 2880 tattgtctct gccctggctagaatgggggc ataatctgag ccttgttccc ttgtccagtg 2940 tggctgaccc ttgacctcttccttcctcct ccctttgttt tgggattcag aaaactgctt 3000 gtcacagaca atttattttttattaaaaaa gatataagct ttaaag 3046 78 1436 PRT Bos sp. 78 Met Ala Leu GlyArg His Leu Ser Leu Arg Gly Leu Cys Val Leu Leu 1 5 10 15 Leu Gly ThrMet Val Gly Gly Gln Ala Leu Glu Leu Arg Leu Lys Asp 20 25 30 Gly Val HisArg Cys Glu Gly Arg Val Glu Val Lys His Gln Gly Glu 35 40 45 Trp Gly ThrVal Asp Gly Tyr Arg Trp Thr Leu Lys Asp Ala Ser Val 50 55 60 Val Cys ArgGln Leu Gly Cys Gly Ala Ala Ile Gly Phe Pro Gly Gly 65 70 75 80 Ala TyrPhe Gly Pro Gly Leu Gly Pro Ile Trp Leu Leu Tyr Thr Ser 85 90 95 Cys GluGly Thr Glu Ser Thr Val Ser Asp Cys Glu His Ser Asn Ile 100 105 110 LysAsp Tyr Arg Asn Asp Gly Tyr Asn His Gly Arg Asp Ala Gly Val 115 120 125Val Cys Ser Gly Phe Val Arg Leu Ala Gly Gly Asp Gly Pro Cys Ser 130 135140 Gly Arg Val Glu Val His Ser Gly Glu Ala Trp Ile Pro Val Ser Asp 145150 155 160 Gly Asn Phe Thr Leu Ala Thr Ala Gln Ile Ile Cys Ala Glu LeuGly 165 170 175 Cys Gly Lys Ala Val Ser Val Leu Gly His Glu Leu Phe ArgGlu Ser 180 185 190 Ser Ala Gln Val Trp Ala Glu Glu Phe Arg Cys Glu GlyGlu Glu Pro 195 200 205 Glu Leu Trp Val Cys Pro Arg Val Pro Cys Pro GlyGly Thr Cys His 210 215 220 His Ser Gly Ser Ala Gln Val Val Cys Ser AlaTyr Ser Glu Val Arg 225 230 235 240 Leu Met Thr Asn Gly Ser Ser Gln CysGlu Gly Gln Val Glu Met Asn 245 250 255 Ile Ser Gly Gln Trp Arg Ala LeuCys Ala Ser His Trp Ser Leu Ala 260 265 270 Asn Ala Asn Val Ile Cys ArgGln Leu Gly Cys Gly Val Ala Ile Ser 275 280 285 Thr Pro Gly Gly Pro HisLeu Val Glu Glu Gly Asp Gln Ile Leu Thr 290 295 300 Ala Arg Phe His CysSer Gly Ala Glu Ser Phe Leu Trp Ser Cys Pro 305 310 315 320 Val Thr AlaLeu Gly Gly Pro Asp Cys Ser His Gly Asn Thr Ala Ser 325 330 335 Val IleCys Ser Gly Asn Gln Ile Gln Val Leu Pro Gln Cys Asn Asp 340 345 350 SerVal Ser Gln Pro Thr Gly Ser Ala Ala Ser Glu Asp Ser Ala Pro 355 360 365Tyr Cys Ser Asp Ser Arg Gln Leu Arg Leu Val Asp Gly Gly Gly Pro 370 375380 Cys Ala Gly Arg Val Glu Ile Leu Asp Gln Gly Ser Trp Gly Thr Ile 385390 395 400 Cys Asp Asp Gly Trp Asp Leu Asp Asp Ala Arg Val Val Cys ArgGln 405 410 415 Leu Gly Cys Gly Glu Ala Leu Asn Ala Thr Gly Ser Ala HisPhe Gly 420 425 430 Ala Gly Ser Gly Pro Ile Trp Leu Asp Asn Leu Asn CysThr Gly Lys 435 440 445 Glu Ser His Val Trp Arg Cys Pro Ser Arg Gly TrpGly Gln His Asn 450 455 460 Cys Arg His Lys Gln Asp Ala Gly Val Ile CysSer Glu Phe Leu Ala 465 470 475 480 Leu Arg Met Val Ser Glu Asp Gln GlnCys Ala Gly Trp Leu Glu Val 485 490 495 Phe Tyr Asn Gly Thr Trp Gly SerVal Cys Arg Asn Pro Met Glu Asp 500 505 510 Ile Thr Val Ser Thr Ile CysArg Gln Leu Gly Cys Gly Asp Ser Gly 515 520 525 Thr Leu Asn Ser Ser ValAla Leu Arg Glu Gly Phe Arg Pro Gln Trp 530 535 540 Val Asp Arg Ile GlnCys Arg Lys Thr Asp Thr Ser Leu Trp Gln Cys 545 550 555 560 Pro Ser AspPro Trp Asn Tyr Asn Ser Cys Ser Pro Lys Glu Glu Ala 565 570 575 Tyr IleTrp Cys Ala Asp Ser Arg Gln Ile Arg Leu Val Asp Gly Gly 580 585 590 GlyArg Cys Ser Gly Arg Val Glu Ile Leu Asp Gln Gly Ser Trp Gly 595 600 605Thr Ile Cys Asp Asp Arg Trp Asp Leu Asp Asp Ala Arg Val Val Cys 610 615620 Lys Gln Leu Gly Cys Gly Glu Ala Leu Asp Ala Thr Val Ser Ser Phe 625630 635 640 Phe Gly Thr Gly Ser Gly Pro Ile Trp Leu Asp Glu Val Asn CysArg 645 650 655 Gly Glu Glu Ser Gln Val Trp Arg Cys Pro Ser Trp Gly TrpArg Gln 660 665 670 His Asn Cys Asn His Gln Glu Asp Ala Gly Val Ile CysSer Gly Phe 675 680 685 Val Arg Leu Ala Gly Gly Asp Gly Pro Cys Ser GlyArg Val Glu Val 690 695 700 His Ser Gly Glu Ala Trp Thr Pro Val Ser AspGly Asn Phe Thr Leu 705 710 715 720 Pro Thr Ala Gln Val Ile Cys Ala GluLeu Gly Cys Gly Lys Ala Val 725 730 735 Ser Val Leu Gly His Met Pro PheArg Glu Ser Asp Gly Gln Val Trp 740 745 750 Ala Glu Glu Phe Arg Cys AspGly Gly Glu Pro Glu Leu Trp Ser Cys 755 760 765 Pro Arg Val Pro Cys ProGly Gly Thr Cys Leu His Ser Gly Ala Ala 770 775 780 Gln Val Val Cys SerVal Tyr Thr Glu Val Gln Leu Met Lys Asn Gly 785 790 795 800 Thr Ser GlnCys Glu Gly Gln Val Glu Met Lys Ile Ser Gly Arg Trp 805 810 815 Arg AlaLeu Cys Ala Ser His Trp Ser Leu Ala Asn Ala Asn Val Val 820 825 830 CysArg Gln Leu Gly Cys Gly Val Ala Ile Ser Thr Pro Arg Gly Pro 835 840 845His Leu Val Glu Gly Gly Asp Gln Ile Ser Thr Ala Gln Phe His Cys 850 855860 Ser Gly Ala Glu Ser Phe Leu Trp Ser Cys Pro Val Thr Ala Leu Gly 865870 875 880 Gly Pro Asp Cys Ser His Gly Asn Thr Ala Ser Val Ile Cys SerGly 885 890 895 Asn His Thr Gln Val Leu Pro Gln Cys Asn Asp Phe Leu SerGln Pro 900 905 910 Ala Gly Ser Ala Ala Ser Glu Glu Ser Ser Pro Tyr CysSer Asp Ser 915 920 925 Arg Gln Leu Arg Leu Val Asp Gly Gly Gly Pro CysGly Gly Arg Val 930 935 940 Glu Ile Leu Asp Gln Gly Ser Trp Gly Thr IleCys Asp Asp Asp Trp 945 950 955 960 Asp Leu Asp Asp Ala Arg Val Val CysArg Gln Leu Gly Cys Gly Glu 965 970 975 Ala Leu Asn Ala Thr Gly Ser AlaHis Phe Gly Ala Gly Ser Gly Pro 980 985 990 Ile Trp Leu Asp Asp Leu AsnCys Thr Gly Lys Glu Ser His Val Trp 995 1000 1005 Arg Cys Pro Ser ArgGly Trp Gly Arg His Asp Cys Arg His Lys Glu 1010 1015 1020 Asp Ala GlyVal Ile Cys Ser Glu Phe Leu Ala Leu Arg Met Val Ser 1025 1030 1035 1040Glu Asp Gln Gln Cys Ala Gly Trp Leu Glu Val Phe Tyr Asn Gly Thr 10451050 1055 Trp Gly Ser Val Cys Arg Ser Pro Met Glu Asp Ile Thr Val SerVal 1060 1065 1070 Ile Cys Arg Gln Leu Gly Cys Gly Asp Ser Gly Ser LeuAsn Thr Ser 1075 1080 1085 Val Gly Leu Arg Glu Gly Ser Arg Pro Arg TrpVal Asp Leu Ile Gln 1090 1095 1100 Cys Arg Lys Met Asp Thr Ser Leu TrpGln Cys Pro Ser Gly Pro Trp 1105 1110 1115 1120 Lys Tyr Ser Ser Cys SerPro Lys Glu Glu Ala Tyr Ile Ser Cys Glu 1125 1130 1135 Gly Arg Arg ProLys Ser Cys Pro Thr Ala Ala Ala Cys Thr Asp Arg 1140 1145 1150 Glu LysLeu Arg Leu Arg Gly Gly Asp Ser Glu Cys Ser Gly Arg Val 1155 1160 1165Glu Val Trp His Asn Gly Ser Trp Gly Thr Val Cys Asp Asp Ser Trp 11701175 1180 Ser Leu Ala Glu Ala Glu Val Val Cys Gln Gln Leu Gly Cys GlyGln 1185 1190 1195 1200 Ala Leu Glu Ala Val Arg Ser Ala Ala Phe Gly ProGly Asn Gly Ser 1205 1210 1215 Ile Trp Leu Asp Glu Val Gln Cys Gly GlyArg Glu Ser Ser Leu Trp 1220 1225 1230 Asp Cys Val Ala Glu Pro Trp GlyGln Ser Asp Cys Lys His Glu Glu 1235 1240 1245 Asp Ala Gly Val Arg CysSer Gly Val Arg Thr Thr Leu Pro Thr Thr 1250 1255 1260 Thr Ala Gly ThrArg Thr Thr Ser Asn Ser Leu Pro Gly Ile Phe Ser 1265 1270 1275 1280 LeuPro Gly Val Leu Cys Leu Ile Leu Gly Ser Leu Leu Phe Leu Val 1285 12901295 Leu Val Ile Leu Val Thr Gln Leu Leu Arg Trp Arg Ala Glu Arg Arg1300 1305 1310 Ala Leu Ser Ser Tyr Glu Asp Ala Leu Ala Glu Ala Val TyrGlu Glu 1315 1320 1325 Leu Asp Tyr Leu Leu Thr Gln Lys Glu Gly Leu GlySer Pro Asp Gln 1330 1335 1340 Met Thr Asp Val Pro Asp Glu Asn Tyr AspAsp Ala Glu Glu Val Pro 1345 1350 1355 1360 Val Pro Gly Thr Pro Ser ProSer Gln Gly Asn Glu Glu Glu Val Pro 1365 1370 1375 Pro Glu Lys Glu AspGly Val Arg Ser Ser Gln Thr Gly Ser Phe Leu 1380 1385 1390 Asn Phe SerArg Glu Ala Ala Asn Pro Gly Glu Gly Glu Glu Ser Phe 1395 1400 1405 TrpLeu Leu Gln Gly Lys Lys Gly Asp Ala Gly Tyr Asp Asp Val Glu 1410 14151420 Leu Ser Ala Leu Gly Thr Ser Pro Val Thr Phe Ser 1425 1430 1435 794308 DNA Bos sp. 79 atggctctgg gcagacacct ctccctgcgg ggactctgtgtcctcctcct cggcaccatg 60 gtgggtggtc aagctctgga gctgaggttg aaggatggagtccatcgctg tgaggggaga 120 gtggaagtga agcaccaagg agaatggggc acagtggatggttacaggtg gacattgaag 180 gatgcatctg tagtgtgcag acagctgggg tgtggagctgccattggttt tcctggaggg 240 gcttattttg ggccaggact tggccccatt tggcttttgtatacttcatg tgaagggaca 300 gagtcaactg tcagtgactg tgagcattct aatattaaagactatcgtaa tgatggctat 360 aatcatggtc gggatgctgg agtagtctgc tcaggatttgtgcgtctggc tggaggggat 420 ggaccctgct cagggcgagt agaagtgcat tctggagaagcttggatccc agtgtctgat 480 gggaacttca cacttgccac tgcccagatc atctgtgcagagttgggttg tggcaaggct 540 gtgtctgtcc tgggacatga gctcttcaga gagtccagtgcccaggtctg ggctgaagag 600 ttcaggtgtg agggggagga gcctgagctc tgggtctgccccagagtgcc ctgtccaggg 660 ggcacgtgtc accacagtgg atctgctcag gttgtttgttcagcatactc agaagtccgg 720 ctcatgacaa acggctcctc tcagtgtgaa gggcaggtggagatgaacat ttctggacaa 780 tggagagcgc tctgtgcctc ccactggagt ctggccaatgccaatgttat ctgtcgtcag 840 ctcggctgtg gagttgccat ctccaccccc ggaggaccacacttggtgga agaaggtgat 900 cagatcctaa cagcccgatt tcactgctct ggggctgagtccttcctgtg gagttgtcct 960 gtgactgccc tgggtggtcc tgactgttcc catggcaacacagcctctgt gatctgctca 1020 ggaaaccaga tccaggtgct tccccagtgc aacgactccgtgtctcaacc tacaggctct 1080 gcggcctcag aggacagcgc cccctactgc tcagacagcaggcagctccg cctggtggac 1140 gggggcggtc cctgcgccgg gagagtggag atccttgaccagggctcctg gggcaccatc 1200 tgtgatgacg gctgggacct ggacgatgcc cgcgtggtgtgcaggcagct gggctgtgga 1260 gaagccctca atgccacggg gtctgctcac ttcggggcaggatcagggcc catctggttg 1320 gacaacttga actgcacagg aaaggagtcc cacgtgtggaggtgcccttc ccggggctgg 1380 gggcagcaca actgcagaca caagcaggac gcgggggtcatctgctcaga gttcctggcc 1440 ctcaggatgg tgagtgagga ccagcagtgt gctgggtggctggaagtttt ctacaatggg 1500 acctggggca gtgtctgccg taaccccatg gaagacatcactgtgtccac gatctgcaga 1560 cagcttggct gtggggacag tggaaccctc aactcttctgttgctcttag agaaggtttt 1620 aggccacagt gggtggatag aatccagtgt cggaaaactgacacctctct ctggcagtgt 1680 ccttctgacc cttggaatta caactcatgc tctccaaaggaggaagccta tatctggtgt 1740 gcagacagca gacagatccg cctggtggat ggaggtggtcgctgctctgg gagagtggag 1800 atccttgacc agggctcctg gggcaccatc tgtgatgaccgctgggacct ggacgatgcc 1860 cgtgtggtgt gcaagcagct gggctgtgga gaagccctggacgccactgt ctcttccttc 1920 ttcgggacgg gatcagggcc catctggctg gatgaagtgaactgcagagg agaggagtcc 1980 caagtatgga ggtgcccttc ctggggatgg cggcaacacaactgcaatca tcaagaagat 2040 gcaggagtca tctgctcagg atttgtgcgt ctggctggaggagatggacc ctgctcaggg 2100 cgagtagaag tgcattctgg agaagcctgg accccagtgtctgatggaaa cttcacactc 2160 cccactgccc aggtcatctg tgcagagctg ggatgtggcaaggctgtgtc tgtcctggga 2220 cacatgccat tcagagagtc cgatggccag gtctgggctgaagagttcag gtgtgatggg 2280 ggggagcctg agctctggtc ctgccccaga gtgccctgtccaggaggcac atgtctccac 2340 agtggagctg ctcaggttgt ctgttcagtg tacacagaagtccagcttat gaaaaacggc 2400 acctctcaat gtgaggggca ggtggagatg aagatctctggacgatggag agcgctctgt 2460 gcctcccact ggagtctggc caatgccaat gttgtctgtcgtcagctcgg ctgtggagtc 2520 gccatctcca cccccagagg accacacttg gtggaaggaggtgatcagat ctcaacagcc 2580 caatttcact gctcaggggc tgagtccttc ctgtggagttgtcctgtgac tgccttgggt 2640 gggcctgact gttcccatgg caacacagcc tctgtgatctgctcaggaaa ccacacccag 2700 gtgctgcccc agtgcaacga cttcctgtct caacctgcaggctctgcggc ctcagaggag 2760 agttctccct actgctcaga cagcaggcag ctccgcctggtggacggggg cggtccctgc 2820 ggcgggagag tggagatcct tgaccagggc tcctggggcaccatctgtga tgatgactgg 2880 gacctggacg atgcccgtgt ggtgtgcagg cagctgggctgtggagaagc cctcaatgcc 2940 acggggtctg ctcacttcgg ggcaggatca gggcccatctggctggacga cctgaactgc 3000 acaggaaagg agtcccacgt gtggaggtgc ccttcccggggctgggggcg gcacgactgc 3060 agacacaagg aggacgccgg ggtcatctgc tcagagttcctggccctcag gatggtgagc 3120 gaggaccagc agtgtgctgg gtggctggag gttttctacaacgggacctg gggcagtgtc 3180 tgccgcagcc ccatggaaga tatcactgtg tccgtgatctgcagacagct tggatgtggg 3240 gacagtggaa gtctcaacac ctctgttggt ctcagggaaggttctagacc ccggtgggta 3300 gatttaattc agtgtcggaa aatggatacc tctctctggcagtgtccttc tggcccatgg 3360 aaatacagtt catgctctcc aaaggaggaa gcctacatctcatgtgaagg aagaagaccc 3420 aagagctgtc caactgctgc cgcctgcaca gacagagagaagctccgcct caggggagga 3480 gacagcgagt gctcagggcg ggtggaggtg tggcacaacggctcctgggg caccgtgtgc 3540 gatgactcct ggagcctggc agaggctgag gtggtgtgtcagcagctggg ctgtggccag 3600 gccctggaag ccgtgcggtc tgcagcattt ggccctggaaatgggagcat ctggctggac 3660 gaggtgcagt gcgggggccg ggagtcctcc ctgtgggactgtgttgcgga gccctggggg 3720 cagagcgact gcaagcacga ggaggatgct ggtgtgaggtgctctggtgt aaggacaaca 3780 ttgcccacga ccacagcagg gaccagaaca acctcaaattctctccctgg catcttctcc 3840 ctgcctgggg ttctctgcct tatcctgggg tcgcttctcttcctggtcct cgtcatcctg 3900 gtgactcagc tactcagatg gagagcagag cgcagagccttatccagcta tgaagatgct 3960 cttgctgaag ctgtgtatga ggagctcgat taccttctgacacagaagga aggtctgggc 4020 agcccagatc agatgactga tgtccctgat gaaaattatgatgatgctga agaagtacca 4080 gtgcctggaa ctccttctcc ctctcagggg aatgaggaggaagtgccccc agagaaggag 4140 gacggggtga ggtcctctca gacaggctct ttcctgaacttctccagaga ggcagctaat 4200 cctggggaag gagaagagag cttctggctg ctccaggggaagaaagggga tgctgggtat 4260 gatgatgttg aactcagtgc cctgggaaca tccccagtgactttctcg 4308

What is claimed is:
 1. An isolated nucleic acid molecule selected fromthe group consisting of: a) a nucleic acid molecule having a nucleotidesequence which is at least 90% identical to the nucleotide sequence ofany of SEQ ID NOs: 45, 46, the nucleotide sequence of a cDNA of a clonedeposited as ATCC® 207220, or a complement thereof; b) a nucleic acidmolecule comprising at least 100 nucleotide residues and having anucleotide sequence identical to at least 100 consecutive nucleotideresidues of any of SEQ ID NOs: 45, 46, and the nucleotide sequence of acDNA of a clone deposited as ATCC® 207220, or a complement thereof; c) anucleic acid molecule which encodes a polypeptide comprising the aminoacid sequence of any of SEQ ID NOs: 47-52, and the amino acid sequenceencoded by a cDNA of a clone deposited as ATCC® 207220, or a complementthereof; d) a nucleic acid molecule which encodes a fragment of apolypeptide comprising the amino acid sequence of any of SEQ ID NOs:47-52 or the amino acid sequence encoded by a cDNA of a clone depositedas ATCC® 207220, wherein the fragment comprises at least 25 consecutiveamino acid residues of any of SEQ ID NOs: 47-52 or the amino acidsequence encoded by a cDNA of a clone deposited as ATCC® 207220; and e)a nucleic acid molecule which encodes a naturally occurring allelicvariant of a polypeptide comprising the amino acid sequence of any ofSEQ ID NOs: 47-52, wherein the nucleic acid molecule hybridizes with anucleic acid molecule consisting of the nucleotide sequence of any ofSEQ ID NOs: 45, 46, and the nucleotide sequence of a cDNA of a clonedeposited as ATCC® 207220, or a complement thereof under stringentconditions.
 2. The isolated nucleic acid molecule of claim 1, which isselected from the group consisting of: a) a nucleic acid having thenucleotide sequence of any of SEQ ID NOs: 45, 46, and the nucleotidesequence of a cDNA of a clone deposited as ATCC® 207220, or a complementthereof; and b) a nucleic acid molecule which encodes a polypeptidehaving the amino acid sequence of any of SEQ ID NOs: 47-52 or the aminoacid sequence encoded by a cDNA of a clone deposited as ATCC® 207220, ora complement thereof.
 3. The nucleic acid molecule of claim 1, furthercomprising vector nucleic acid sequences.
 4. The nucleic acid moleculeof claim 1 further comprising nucleic acid sequences encoding aheterologous polypeptide.
 5. A host cell which contains the nucleic acidmolecule of claim
 1. 6. The host cell of claim 5 which is a mammalianhost cell.
 7. A non-human mammalian host cell containing the nucleicacid molecule of claim
 1. 8. An isolated polypeptide selected from thegroup consisting of: a) a fragment of a polypeptide comprising the aminoacid sequence of any of SEQ ID NOs: 47-52 or the amino acid sequenceencoded by a cDNA of a clone deposited as ATCC® 207220, wherein thefragment comprises at least 25 contiguous amino acids of any of SEQ IDNOs: 47-52 or the amino acid sequence encoded by a cDNA of a clonedeposited as ATCC® 207220; b) a naturally occurring allelic variant of apolypeptide comprising the amino acid sequence of any of SEQ ID NOs:47-52 or the amino acid sequence encoded by a cDNA of a clone depositedas ATCC® 207220, wherein the polypeptide is encoded by a nucleic acidmolecule which hybridizes to a nucleic acid molecule consisting of thenucleotide sequence of any of SEQ ID NOs: 45, 46, and the nucleotidesequence of a cDNA of a clone deposited as ATCC® 207220, or a complementthereof under stringent conditions; and c) a polypeptide which isencoded by a nucleic acid molecule comprising a nucleotide sequencewhich is at least 90% identical to a nucleic acid consisting of thenucleotide sequence of any of SEQ ID NOs: 45, 46, and the nucleotidesequence of a cDNA of a clone deposited as ATCC® 207220, or a complementthereof.
 9. The isolated polypeptide of claim 8 having the amino acidsequence of any of SEQ ID NOs: 47-52 or the amino acid sequence encodedby a cDNA of a clone deposited as ATCC® 207220, or a complement thereof.10. The polypeptide of claim 8, wherein the amino acid sequence of thepolypeptide further comprises heterologous amino acid residues.
 11. Anantibody which selectively binds with the polypeptide of claim
 8. 12. Amethod for producing a polypeptide selected from the group consistingof: a) a polypeptide having an amino acid sequence comprising any of SEQID NOs: 47-52 or the amino acid sequence encoded by a cDNA of a clonedeposited as ATCC® 207220, or a complement thereof; b) a polypeptidecomprising a fragment of a protein having the amino acid sequence of anyof SEQ ID NOs: 47-52 or the amino acid sequence encoded by a cDNA of aclone deposited as ATCC® 207220, or a complement thereof, wherein thefragment comprises at least 25 contiguous amino acid residues of any ofSEQ ID NOs: 47-52 or the amino acid sequence encoded by a cDNA of aclone deposited as ATCC® 207220, or a complement thereof; and c) anaturally occurring allelic variant of a polypeptide having an aminoacid sequence comprising the sequence of any of SEQ ID NOs: 47-52 or theamino acid sequence encoded by a cDNA of a clone deposited as ATCC®207220, or a complement thereof, wherein the polypeptide is encoded by anucleic acid molecule which hybridizes with a nucleic acid moleculeconsisting of the nucleotide sequence of any of SEQ ID NOs: 45, 46, andthe nucleotide sequence of a cDNA of a clone deposited as ATCC® 207220,or a complement thereof under stringent conditions, the methodcomprising culturing the host cell of claim 5 under conditions in whichthe nucleic acid molecule is expressed.
 13. A method for detecting thepresence of a polypeptide of claim 8 in a sample, comprising: a)contacting the sample with a compound which selectively binds with apolypeptide of claim 8; and b) determining whether the compound bindswith the polypeptide in the sample.
 14. The method of claim 13, whereinthe compound which binds with the polypeptide is an antibody.
 15. A kitcomprising a compound which selectively binds with a polypeptide ofclaim 8 and instructions for use.
 16. A method for detecting thepresence of a nucleic acid molecule of claim 1 in a sample, comprisingthe steps of: a) contacting the sample with a nucleic acid probe orprimer which selectively hybridizes with the nucleic acid molecule; andb) determining whether the nucleic acid probe or primer binds with anucleic acid molecule in the sample.
 17. The method of claim 16, whereinthe sample comprises mRNA molecules and is contacted with a nucleic acidprobe.
 18. A kit comprising a compound which selectively hybridizes witha nucleic acid molecule of claim 1 and instructions for use.
 19. Amethod for identifying a compound which binds with a polypeptide ofclaim 8, the method comprising the steps of: a) contacting apolypeptide, or a cell expressing a polypeptide of claim 8 with a testcompound; and b) determining whether the polypeptide binds with the testcompound.
 20. The method of claim 19, wherein the binding of the testcompound with the polypeptide is detected by a method selected from thegroup consisting of: a) detection of binding by direct detecting of testcompound/polypeptide binding; b) detection of binding using acompetition binding assay; and c) detection of binding using an assayfor an activity characteristic of the polypeptide.
 21. A method formodulating the activity of a polypeptide of claim 8 comprisingcontacting the polypeptide or a cell expressing the polypeptide with acompound which binds with the polypeptide in a sufficient concentrationto modulate the activity of the polypeptide.
 22. A method foridentifying a compound which modulates the activity of a polypeptide ofclaim 8, comprising: a) contacting the polypeptide with a test compound;and b) determining the effect of the test compound on the activity ofthe polypeptide to thereby identify a compound which modulates theactivity of the polypeptide.
 23. An antibody substance which selectivelybinds to the polypeptide of claim 8, wherein the antibody substance ismade by providing the polypeptide to an immunocompetent vertebrate andthereafter harvesting blood or serum from the vertebrate.
 24. A methodof assessing whether a first human patient is afflicted with anepithelial or endothelial tumor, the method comprising comparing: a)occurrence of a nucleic acid molecule of claim 1 in a sample obtainedfrom the first patient and b) occurrence of the nucleic acid molecule ina control sample selected from the group consisting of i) a controlsample obtained from a tissue that is obtained from the first patientand that is known not to comprise the tumor; and ii) a control sampleobtained from a second patient who is known not to be afflicted with thetumor, whereby a difference between the first sample and the controlsample is an indication that the patient is afflicted with the tumor.25. The method of claim 24, wherein the tumor is selected from the groupconsisting of a colon tumor, a prostate tumor, a lung tumor, apancreatic tumor, and a breast tumor.
 26. The method of claim 25,wherein the tumor is a colon tumor.
 27. A method of assessing whether afirst human patient is afflicted with an epithelial or endothelialtumor, the method comprising comparing a) occurrence of a nucleic acidmolecule of claim 1 in a sample obtained from the first patient and b)occurrence of the nucleic acid molecule in a control sample selectedfrom the group consisting of i) a control sample obtained from a tissuethat is obtained from the first patient and that is known to comprisethe tumor; and ii) a control sample obtained from a second patient whois known to be afflicted with the tumor, whereby a difference betweenthe first sample and the control sample is an indication that thepatient is not afflicted with the tumor.
 28. A method of screening foragents which decrease the activity of a TANGO-294-like lipase protein,the method comprising: contacting a test compound with a TANGO 294-likelipase polypeptide encoded by an isolated nucleic acid molecule of claim1 and detecting binding between the test compound and the TANGO 294-likelipase polypeptide, wherein binding between the test compound and theTANGO 294-like lipase polypeptide is an indication that the testcompound is an agent which decreases the activity of the TANGO 294-likelipase protein.
 29. A method of screening for agents which modulate theactivity of a TANGO 294-like lipase protein, the method comprising:contacting a test compound with a TANGO 294-like lipase polypeptideencoded by an isolated nucleic acid molecule of claim 1 and detecting aTANGO 294-like lipase activity of the polypeptide, wherein increasedTANGO 294-like lipase activity in the presence of the test compound isan indication that the test compound is an agent useful for increasingthe activity of the TANGO 294-like lipase protein, and decreased TANGO294-like lipase activity in the presence of the test compound is anindication that the test compound is an agent useful for decreasing theactivity of the TANGO 294-like lipase protein.
 30. A method of screeningfor agents which decrease the activity of a TANGO 294-like lipaseprotein, the method comprising: contacting a test compound with anisolated nucleic acid molecule of claim 1 and detecting binding of thetest compound with the isolated nucleic acid molecule, wherein bindingbetween the test compound and the isolated nucleic acid molecule is anindication that the test compound is a agent useful for decreasing theactivity of the TANGO 294-like lipase protein.
 31. A method of reducingthe activity of a TANGO 294-like lipase protein of a cell, the methodcomprising contacting the cell with a reagent which specifically bindswith an isolated nucleic acid molecule of claim 1, whereby the activityof the TANGO 294-like lipase protein is reduced.
 32. A method ofreducing the activity of a TANGO 294-like lipase protein of a cell, themethod comprising contacting the cell with a reagent which specificallybinds with an isolated polypeptide of claim 8, whereby the activity ofthe TANGO 294-like lipase protein is reduced.
 33. A method of making apharmaceutical composition, the method comprising identifying an agentaccording to the method of claim 28 and combining the agent and apharmaceutically acceptable carrier to form the pharmaceuticalcomposition.
 34. A method of modulating the activity of a TANGO 294-likelipase protein in a TANGO 294-related disorder, the method comprisingmaking the pharmaceutical composition according to claim 33 andadministering the pharmaceutical composition to a human afflicted withthe disorder.
 35. The method of claim 34, wherein the disorder isselected from the group consisting of a tumor, a disorder of fatabsorption, a disorder of fat metabolism, a blood flow disorder, a bloodpressure disorder, an inflammatory disorder, an immune disorder, athrombotic disorder, and a disorder involving inappropriate plateletadherence.
 36. The method of claim 35, wherein the disorder is a tumorof endothelial or epithelial origin.
 37. The method of claim 35, whereinthe disorder is a colon tumor.
 38. The method of claim 35, wherein thedisorder is a pancreatic tumor.
 39. The method of claim 35, wherein thedisorder is selected from the group consisting of inadequate expressionof gastric lipase, inadequate expression of pancreatic lipase, cysticfibrosis, exocrine pancreatic insufficiency, and obesity.
 40. The methodof claim 35, wherein the disorder is selected from the group consistingof arterial hypertension, renovascular hypertension, syncope,orthostatic hypotension, and shock.
 41. The method of claim 35, whereinthe disorder is an inflammatory disorder selected from the groupconsisting of gastritis, gastric ulcer, colitis, irritable bowelsyndrome, inflammatory bowel syndrome, dermatitis, and pancreatitis. 42.The method of claim 35, wherein the disorder is an autoimmune disorderselected from the group consisting of rheumatoid arthritis, psoriasis,myasthenia gravis, an allergy, insulin resistance, systemic lupuserythematosus, scleroderma, and autoimmune diabetes mellitus.
 43. Themethod of claim 35, wherein the disorder is an infection of a human byan infectious agent.
 44. The method of claim 43, wherein the infectiousagent is human immunodeficiency virus.
 45. The method of claim 35,wherein the disorder is selected from the group consisting ofhemophilia, stroke, myocardial infarction, coronary artery disease, andatherosclerosis.
 46. A method of making a pharmaceutical composition,the method comprising identifying an agent according to the method ofclaim 29 and combining the agent and a pharmaceutically acceptablecarrier to form the pharmaceutical composition.
 47. A method ofmodulating the activity of a TANGO 294-like lipase protein in a TANGO294-related disorder, the method comprising making the pharmaceuticalcomposition according to claim 46 and administering the pharmaceuticalcomposition to a human afflicted with the disorder.
 48. The method ofclaim 47, wherein the disorder is selected from the group consisting ofa tumor, a disorder of fat absorption, a disorder of fat metabolism, ablood flow disorder, a blood pressure disorder, an inflammatorydisorder, an immune disorder, a thrombotic disorder, and a disorderinvolving inappropriate platelet adherence.
 49. A method of making apharmaceutical composition, the method comprising identifying an agentaccording to the method of claim 30 and combining the agent and apharmaceutically acceptable carrier to form the pharmaceuticalcomposition.
 50. A method of modulating the activity of a TANGO 294-likelipase protein in a TANGO 294-related disorder, the method comprisingmaking the pharmaceutical composition according to claim 49 andadministering the pharmaceutical composition to a human afflicted withthe disorder.
 51. The method of claim 50, wherein the disorder isselected from the group consisting of a tumor, a disorder of fatabsorption, a disorder of fat metabolism, a blood flow disorder, a bloodpressure disorder, an inflammatory disorder, an immune disorder, athrombotic disorder, and a disorder involving inappropriate plateletadherence.